Phage display

Definition

Phage display is a laboratory platform that allows scientists to study protein interactions on a large-scale and select proteins with the highest affinity for specific targets.

The photograph shows a phage as seen under a microscope. Credit: Ian Hands-Portman.

Importance

The key advantage of phage display is that it provides a means to identify target-binding proteins from a library of millions of different proteins without the need to screen each molecule individually. This makes it possible to screen billions of proteins each week. By linking a selected protein with its encoding gene, phage display also provides a means to easily identify coding sequences of binding proteins. These can be stored, amplified or processed in other ways. Phage display is a pivotal tool for early basic scientific research and for the development of new drugs and vaccines. The technology has proved particularly important to the production of safer and more effective monoclonal antibody drugs. Phage display libraries consisting entirely of human antibody sequences, for example, have made it possible to produce fully human antibodies. The first fully human therapeutic antibody (adalimumab), a blockbuster drug, generated US$9.3 billion in annual sales in 2012. Four fully human therapeutic antibodies currently approved as treatments in the US and the UK were developed using phage display and many more are in the pipeline. Phage display is also used to develop vaccines for conditions such as prostate cancer and HIV. It is also an important tool for the generation of diagnostic tests for monitoring disease progression and evaluating treatment efficacy.

Discovery

Phage display was first described by George P Smith in 1985, who deployed it as a method to identify a gene against which he had raised antibodies. The technique was taken further by Greg Winter and John McCafferty at the Laboratory of Molecular Biology in Cambridge, UK, and Richard Lerner and Carlos F. Barbas at The Scripps Research institute, US, who independently used phage display to build large libraries of fully human antibody sequences. This work laid the foundation for the development of human antibody based drugs. Today there are several types of phage display libraries, including peptide libraries, protein libraries and antibody libraries.

Application

The phage display technique relies on the generation of a library of millions of bacteriophages that have been genetically engineered to display different peptides or proteins on their surface. This is achieved by inserting a gene encoding a protein of interest into the phage's protein shell, which sets up a direct physical link between DNA sequences and their encoding proteins. The aim of the modification is to generate a molecule that can mimic a natural modulator within the cellular process. Several types of phages are used for the purpose of phage display. Filamentous bacteriophages are the most popular.

The genetically modified phages are assembled into a library for use as a platform to screen proteins, peptides and DNA sequences. Screening is performed by the addition of the phage-display library to the wells of a microtiter plate that contains immobilised target proteins or DNA sequences. The plate is then incubated for some time to allow the phages to bind with the target of interest and then washed so as to flush away any non-binding phages. Any phages that remain attached to the wells are then removed and inserted into other bacteria for replication. The cycle is repeated until only phage-displaying proteins highly specific for the target remain. Once the whole process is completed, the gene coding for the specific protein is isolated and purified from the phage so that it can be used for different applications.

Any laboratory that is well versed in basic microbiology can easily make a phage display library with minimum expense. This has allowed phages to become an invaluable tool for basic immunological and cellular biological research as well as for the development of new drugs and vaccines. Prior to the development of phage display, drug development depended on the time-consuming and expensive process of combing through hundreds of thousands of components using tests conducted in test tubes or laboratory dishes.

This section was put together by Lara Marks with Gavin Hubbard.

Phage display: timeline of key events

d'Herelle was a microbiologist who co-discovered bacteriophages (phages), viruses that infect bacteria that are now major tools in biotechnology. He isolated the first phage from chicken faeces in 1919. Following this he successfully treated chicken affected by a plague of typhus with the phage and in August 1919 cured a patient with dysentery using the same method. This laid the basis for the development of phage therapy. 1873-04-25T00:00:00+0000Twort was a bacteriologist who in 1915 discovered the first virus (bacteriophage) that infects bacteria. He found the virus after experiencing difficulties growing the varrina virus, a key component of the smallpox vaccine, on agar plates. Unable to take his work further because of the war, Twort's discovery was largely ignored at the time. Today bacteriophages are a vital tool in biotechnology and underpin the development of phage therapy, a treatment gaining popularity for the treatment of bacterial infections. 1877-10-22T00:00:00+0000An English bacteriologist, Ernest Hanbury Hankin, described a substance taken from the Ganga and Jumma rivers in India, that could destroy cholera-inducing bacteria. He found the substance to be smaller than bacteria. E.H. Hankin, 'L’action "bactéricide des eaux de la Jumna et du Gange sur le vibrion du cholera', Annals Institute Pasteur, 10 (1896) 511-23.1896-01-01T00:00:00+0000Term 'bacteriophage, was coined by Felix d'Herelle in 1917. 1915-01-01T00:00:00+0000Bacteriophages, also called phages, are a group of viruses that infect bacteria. It was first observed by the English bacteriologist Frederick Twort who published his finding in 'An Investigation on the Nature of Ultra-Microscopic Viruses', The Lancet. 186/4814 (1915), 1241–43, doi:10.1016/S0140-6736(01)20383-3. Twort's finding was initially ignored until after Félix d'Herelle independently also noted phages in 1917. Phages are now an important tool in molecular biology. 1915-12-04T00:00:00+0000Frederick W Twort, an English bacteriologist, noticed a filterable transparent material that could entirely break down bacteria when trying to grow the vaccina virus. F.W. Twort, 'An investigation on the nature of ultra-microscopic viruses', The Lancet, 2 (1915), 1241-3). Financial difficulties and war prevented Twort from pursuing the matter further.1915-12-04T00:00:00+0000The term was coined by Felix d'Herelle, a French-Canadian microbiologist, after he noticed anti-Shigella microbes in the filtrate of dysentery fluids taken from patients recovering from shigellosis F. d'Herelle, 'Sur un microbe invisible antagoniste des bacilles dysentériques', Comptes rendus de l'Académie des Sciences', 165 (1917), 373-5.1917-09-10T00:00:00+0000D’Herelle treated four patients (three of whom were brothers, aged 3, 7, 12) who suffered from severe dysentery in Paris under the clinical supervision of Victor-Henri Hutinel. This study was published later in 1931.1919-01-01T00:00:00+0000Winner of the Nobel Prize for his work on immunity, Bordet believed phages were a lytic enzyme.1919-01-01T00:00:00+0000Lederberg is best known for having discovered the lambda phage, an indispensable tool for studying gene regulation and genetic recombination. She also invented the replica plating technique which is pivotal to tracking antibiotic resistance. 1922-12-18T00:00:00+0000George Eliava learnt about bacteriophage from d'Herelle and became fascinated with the idea while he was working at the Pasteur Institute in the late 1910s. He founded the Eliava Institute in Tbilisi to further study phage therapy.1923-01-01T00:00:00+0000Jules Bordet and Oskar Bail showed phages to have two reproductive patterns: the lysogenic and lytic cycles. In the case of lysogeny, the phage's nucleic acid is inserted into the bacterial host's genome. The inserted phage's DNA, called a prophage, then gets replicated alongside the host's normal reproduction cycle. By contrast in the lytic cycle the the virus replicates separately from the host bacterial DNA and eventually triggers a cell lysis to release its progeny. O. Bail, Med. Klin. (Munich) 21 (1925), 1271–73; J. Bordet, Ann. Inst. Pasteur, 39 (2005), 711–63.1925-01-01T00:00:00+0000D'Herelle treated four Egyptian patients in Alexandira suffering from Y.pestis infections. He developed the treatment based on a highly virulent anti-plague phage that been isolated from rat faeces in Indo-China in 1920. The patients made a full recovery after a month. 1925-01-01T00:00:00+0000D'Herelle supplied some plague phages to Haffking Institute in Bombay, India. The phages initially did not work because they could not be grown in the usual growth medium, made from a digest of macerated pigs, which offended both Muslim and Hindus. D'Herelle took unpaid leave to resolve the issue. He managed to cultivate the phages by using goat tissue, dissolved it in papaya juice. This provided a stock of active anti-plague phages for treating patients. 1926-01-01T00:00:00+0000The Inquiry was organised by the government of British India, the Indian Research Fund Association, several hospitals and research institutes.1927-01-01T00:00:00+0000Conducted by the Bacteriophage Inquiry, the study compared treatment with a control group which included patients who refused treatment. Overall the mortality rate of the untreated group was 62% compared to the phage treated group which was 8%. 1927-07-01T00:00:00+0000D'Herelle took up a position at Yale University.1927-08-01T00:00:00+0000The work was undertaken by d'Herelle, Reginald Malone and Dr. M.N. Lahiri under the umbrella of the Bacteriophage Inquiry. Results from the study was reported in the Indian Medical Gazette in November 1927. 1927-11-01T00:00:00+0000Compiled from the Bacteriophage Inquiry, d'Herelle's results were considered of 'particular importance' by the Congress which recommended further studies in the area.1927-12-01T00:00:00+0000Carried out by J. Morison from King Edward VII Pasteur Institute, India, under the auspices of the Bacteriophage Inquiry, the study suffered from poor staff cooperation.1928-01-01T00:00:00+0000"Eugene Wollman put forward the theory that bacteriophages could transmit certain traits like lysis genetically to their descendents. E. Wollman, ‘Bacteriophage et processus similaire: Hérédité ou Infection?’ Bulletin Institute Pasteur, Paris, 26 (1928), 1-14. "1928-01-01T00:00:00+0000Carried out on behalf of the Bacteriophage Inquiry, two of the studies showed positive results, but a study in Puri Cholera Hospital proved disappointing. This was attributed to poor staff cooperation, compliance and inaccurate diagnoses.1928-08-01T00:00:00+0000Moisei Mel'nyk and his colleagues initiated a programme of testing phages to treat dysentery in Alchevsk, Rykovo, and Krasnyi Luch hospitals in Stalino (now known as Donetsk). The team made regular expeditions to the Donbass region where they found frequent outbreaks of scarlet fever, typhoid and dysentery. Anti-Shigella bacteriophages were isolated from local waters. The programme involved a total of 282 treated patients with 1059 untreated controls. Mortality was halved in phage patients and their recovery was much quicker.'1929-01-01T00:00:00+0000Asheshov's change in direction resulted in his grant funds being cut from 64,260 to 8,500 rupees.1929-01-01T00:00:00+0000Stahl is a molecular biologist and geneticist who helped to elucidate how DNA is replicated. Together with Matthew Medelsohn, Stahl showed that the double-stranded helix molecule of DNA separates into two strands and that each of these strands serve as a template for the production of a new strand of DNA. They did this in 1958. Following this work, Stahl did extensive work on bacteriophages, viruses that infect bacteria, and their genetic recombination. In 1964 he established that DNA in T4 bacteriophages is circular rather than linear. Eight years later he and his wife, Mary, found a DNA sequence in the lambda bacteriophage necessary to initiate genetic recombination. This laid the foundation for genetic engineering. 1929-10-08T00:00:00+0000D’Herelle used the Parisian courts to force the editor of the Annals of the Pasteur Institute to publish his challenge to Bordet to a scientific duel. Most textbooks at the time sided with Bordet.1931-01-01T00:00:00+0000D'Herelle summarised all of the experiments he had so far carried out with phage therapy. Most of them had shown positive results. His paper, however, mostly cited facts without elaborating on each experiment. The methods of administration varied from case to case. F. D’Herelle, ‘Bacteriophage as a treatment in acute medical and surgical infections’, Bulletin New York Academy of Medicine, 7 (1931), 329-48.1931-05-01T00:00:00+0000The Indian Research Fund Association concluded the Inquiry failed to provide conclusive evidence about the effectiveness of phage therapy for cholera due to the unreliable data obtained under field conditions. 1933-01-01T00:00:00+0000Using serological methods, Frank Macfarlane Burnet showed the existence of different species of phages and that they could be distinguished with regard to the range of bacteria species that they lysed. F.M. Burnet, ’The classification of coli-dysentery bacteriophages. III. A correlation of the serological classification with certain biochemical tests’, Journal of Pathology & Bacteriology, 37/2 (1933), 179–84. 1933-01-01T00:00:00+0000After conflicts over his research directions and money, d’Herelle left his position at Yale University and went to work at the Eliava Institute of Bacteriophages, Microbiology and Virology in Georgia.1933-01-01T00:00:00+0000The report was highly critical of phage therapy because of the poor experimental designs d'Herelle had used to test it out. M.D. Eaton, S Bayne-Jones, ‘Bacteriophage therapy: review of the principles and results of the use of bacteriophage in the treatment of infections’, JAMA, 10323 (1934), 1769–76.1934-01-01T00:00:00+00001934-01-01T00:00:00+00001935-01-01T00:00:00+00001936-01-01T00:00:00+0000Increasing political unrest in India and various difficulties in administering field studies and a sanitary reform, which promised to reduce cholera, contributed to the closing of the Inquiry.1936-01-01T00:00:00+0000Eliava was executed on the pretext of anti-Soviet activities during the Great Terror including spying for France, sabotaging vaccines and poisoning wells with bacteriophages. Much speculation exists about the motivation behind his arrest, including a feud with Lavrenti Beria, chief of the secret police, for competing for a woman. Eliava's name was erased from most records, including his own research institute. His name only resurfaced in the late 1960s.1937-07-10T00:00:00+0000The therapy was administered to the local population and phages were also put into wells and ponds. 1938-01-01T00:00:00+0000The new combined institute,, aka Tbilisi Institute, opened in 1939.1938-01-01T00:00:00+0000A team of 11 people, including surgeons, bacteriologists and laboratory assistants, deployed a phage therapy developed at the Tbilsi Institute to treat the soldiers. They found that the treatment worked best when given within the first few hours of the wound getting infected. Elimination of the infection with phages enabled surgeons to suture the wound a week later. 1939-01-01T00:00:00+00001939-01-01T00:00:00+0000H. Ruska, 'Uber die Sichtbarmachung der bakteriophagen Lyse im Ubermikroskop', Naturwissenschaften, 28 (1940), 45–6; E. Pfankuch, GA Kausche, 'Isolierung und übermikroskopische Abbildung eines Bakteriophagen', Naturwissenschaften, 28 (1940), 46.1940-01-01T00:00:00+0000Ernst Ruska produced the first electron micrograph of a phage in pre-war Germany. It was first published in H. Ruska, 'Uber die Sichtbarmachung der bakteriophagen Lyse im Ubermikroskop', Naturwissenschaften, 28 (1940), 45-6. World War II delayed the distribution of Ruska's phage image. 1940-01-01T00:00:00+0000More than 200,000 litres of phage therapies were produced to treat purulent infections and gangrene.1941-01-01T00:00:00+0000The second AMA report supported Bordet’s concept of phage as an auto-catalytically activated lytic principle. It stated phage 'is a protein of high molecular weight and appears to be formed from a precursor originating within the bacterium'. AP Krueger, EJ Scribner, 'The Bacteriophage', JAMA, 116 (1941), 2160-7, 2269-77.1941-05-10T00:00:00+00001941-06-22T00:00:00+0000Zanida Ermolieve from the Institute of Experimental Medicine established a secret underground laboratory in besieged Leningrad to produce cholera phage therapy. The treatment was given to nearly 50,000 people every day. 1942-01-01T00:00:00+0000SE Luria, TF Anderson, 'The identification and characterization of bacteriophages with the electron microscope', PNAS USA, 28/4 (1942), 127-30.1942-04-01T00:00:00+0000The AMA fully accepted d’Herelle’s concept of phage as a virus based on electron micron image. H.E. Morton, FB Engley, 'Dysentery Bacteriophage', JAMA , 127 (1945), 584-91.1945-01-01T00:00:00+0000d'Herelle was a French Canadian microbiologist who co-discovered bacteriophages (phages), viruses that infect bacteria that are now major tools in biotechnology. He isolated the first phage from chicken faeces in 1919. Following this he successfully treated chicken affected by a plague of typhus with the phage and in August 1919 cured a patient with dysentery using the same method. This laid the basis for the development of phage therapy. 1949-02-22T00:00:00+0000The lambda phage has become a key tool in molecular biology and is important for genetic engineering. It has the advantage that it can be easily grown in E Coli and is not pathogenic except in the case of bacteria. Lederberg's discovery paved the way to understanding the transfer of genetic material between bacteria, the mechanisms involved in gene regulation and how piece of DNA break apart and recombine to make new genes. EM Lederberg, 'Lysogenicity in Escherichia coli strain K-12', Microbial Genetics Bulletin, 1, (1950), 5-9. 1950-01-01T00:00:00+0000Twort was an English bacteriologist who in 1915 discovered the first virus (bacteriophage) that infects bacteria. He found the virus after experiencing difficulties growing the varrina virus, a key component of the smallpox vaccine, on agar plates. Unable to take his work further because of the war, Twort's discovery was largely ignored at the time. Today bacteriophages are a vital tool in biotechnology and underpin the development of phage therapy, a treatment gaining popularity for the treatment of bacterial infections.1950-03-20T00:00:00+0000Founded by Professor Ludwik Hirszfeld, the institute focuses on the isolation, characterization and human application of phages. Between 1981 and 1999, they cured approximately 2000 people with phages, mostly after the failure of antibiotics. The institute has developed phages for the treatment of Shigella, septicemia, furunculosis and pulmonary and urinary tract infections.1952-01-01T00:00:00+0000The study involved 30,769 children (six months – seven years old), with 17,044 of them being treated once a week with oral anti-shigella phages, and the remainder being given a placebo. Those who received the therapy experienced a decrease in dysentery over those given the placebo (From 6.7 to 1.8 incidence per 1000). EG Babalova, KT Katsitadze, LA Sakvaredidze, et al. 'Preventive value of dried dysentery bacteriophage', Zh Mikrobiol Epidemiol Immunobiol, 2(1968), 143–45 (in Russian).1963-01-01T00:00:00+0000The treatment was given to mice, calves, piglets and lambs infected with diarrhoea-causing E.coli. The phage mixture was given either by an intramuscular injection or oral administration. HW Smith, M.B Huggins, Journal General Microbiology, 128 (1982), 307-18; HW Smith, M.B Huggins, Journal General Microbiology, 129 (1983), 2659-75; H.W. Smith, M.B Huggins, Journal General Microbiology, 133 (1987), 1111-26; HW Smith, M.B Huggins, KM Shaw, Journal General Microbiology, 133 (1987), 1127-35.1982-01-01T00:00:00+0000The American scientist George Smith develops a method using bacteriophages for studying protein-protein, protein-peptide and protein-DNA interaction. Today phage display is a major tool for drug discovery.1985-01-01T00:00:00+0000J.S. Soothill, J.C. Lawrence, G.A.J. Ayliffe, 'The efficacy of phages in the prevention of the destruction of pig skin in vitro by Pseudomonas aeruginosa', Medical Science Research, 16 (1988), 1287–88; JS Soothill, 'Treatment of experimental infections of mice by bacteriophage. Journal Medical Microbiology, 37 (1992), 258–61; JS Soothill, 'Bacteriophage prevents destruction of skin grafts by P. aeruginosa', Burns, 20 (1994), 209–11. 1988-01-01T00:00:00+0000Gregory Winter together with David Chiswell set up CAT to develop phage display technology for monoclonal antibodies1989-01-01T00:00:00+0000Gregory Winter together with CAT create the first phage monoclonal antibodies, laying the foundation for the generation of diverse libraries of randomly shaped human antibodies. With this scientists are no longer dependent on the natural immune system of animals or humans and the limitations this poses for the production of monoclonal antibodies. 1990-01-01T00:00:00+0000This is achieved by Richard Lerner and Carlos Barbas at the Scripps Research Institute with the backing of Stratagene, an American biotechnology specialising in antibody engineering.1991-01-01T00:00:00+0000Alexander Sulakvelidze and J. Glenn Morris formed a company, Intralytix, in order to license phage treatment for Vancomycin-resistant Enterococcus. However, they could not get an approval due to the strict criteria of clinical trials and the huge uncertainty over phage therapy.1998-01-01T00:00:00+0000The drug, adalimumab (Humira), was approved by the FDA for the treatment of rheumatoid arthritis. It was created using phage display, a technique first invented by Greg Winter in 1990. Cambridge Antibody Technology, BASF Bioresearch Corporation and Abbott Laboratories partnered to develop and market the drug. 2002-12-31T00:00:00+0000Different life cycles of phages; lytic, lysogenic, chronic infection (lysogenic that mutated and lost lytic property, becoming permanent prophage) and pseudo lysogenic (stalled development with no lysis or multiplication similar to lysogenic cycles) published in MG Weinbauer, 'Ecology of prokaryotic viruses', FEMS Microbiology Review, 28 (2004), 127-81.2004-01-01T00:00:00+0000Organised by the American Society for Microbiology, the summit was the first major international gathering devoted to phage biology with over 350 conferees from 24 countries.2004-08-01T00:00:00+0000Intralytix won FDA approval for its first phage product ListShield (formerly known as LMP-102), a food additive that targets Listeria monocytogenes in meat. The preparation is made from 6 different phages to be used in poultry products as an antimicrobial agent against Listeria. Listeriosis can cause life-threatening diseases such as sepsis and meningitis. L.H. Lang, 'FDA approves use of bacteriophages to be added to meat and poultry products', Gastroenterology, 131/5 (2006), 1370.2006-01-01T00:00:00+0000Lederberg is best known for having discovered the lambda phage, an indispensable tool for studying gene regulation and genetic recombination. She also invented the replica plating technique which is pivotal to tracking antibiotic resistance. 2006-11-11T00:00:00+0000The clinical trial evaluated the safety of phage therapy to treat chronic venous leg ulcers infected with S.aureus and P. aeruginosa. The phages were administered topically and recorded to have no difference in frequency of adverse effects. D.D. Rhoads et al, 'Bacteriophage therapy of venous leg ulcers in humans: Results of a phase I safety trial', Journal Wound Care, 18 (2009), 237-8, 240-3.2009-01-01T00:00:00+0000The first controlled clinical trial of a therapeutic phage preparation showed efficacy and safety in the treatment of a chronic otitis, caused by an antibiotic-resistant P.aeruginosa. 12 out of 24 patients were randomly selected for treatment with a combination of one or more of the six phages present in Biophage-PA. The phage treated group had a significantly lower bacterial count after 42 days. A. Wright, C.H. Hawkins, E.E. Anggard, D.R. Harper, 'A controlled clinical trial of a therapeutic bacteriophage preparation in chronic otitis due to antibiotic-resistant pseudomonas aeruginosa; a preliminary report of efficacy', Clinical Otolaryngology, 34/4 (2009), 349-57.2009-01-01T00:00:00+0000Intralytix developed the 'ShigaShield' phage cocktail after failing to secure a license for a phage cocktail for human therapy. The cocktail is active against S. flexneri, S. sonnei and S. dysenteriae, bacteria linked to food poisoning. 2016-01-01T00:00:00+0000AmpliPhi Biosciences, now incorporated into a joint company, Armata, announced the dosing of the first patient in phase 1 clinical trial of AB-SA01.2016-01-01T00:00:00+0000The patient, Tom Patterson, was successfully given intravenous bacteriophage therapy to treat his necrotizing pancreatitis which had the complication of a multidrug resistant A. baumannii infection for which there was no effective antibiotic treatment. R.T. Schooley, B. Biswas, J.J. Gill, et al, 'Development and use of personalized bacteriophage-based therapeutic cocktails to treat a patient with a disseminated resistant A. Baumannii infection'. Antimicrobial Agents and Chemotherapy 61/10 (2017).2017-10-01T00:00:00+0000
Date Event People Places
25 Apr 1873Felix d'Herelle was born in Montreal, Canadad'HerellePasteur Institute
22 Oct 1877Frederick W Twort was born in Camberley, Surrey, United KingdomTwortBrown Animal Sanatory Institution
1896First observation of substance with potential antibacterial activityHankinPasteur Institute
1915Discovery of bacteriophages, type of virus that attacks bacteria, by English bacteriologist William TwortTwortUniversity of London
4 Dec 1915First bacteriophage discoveredTwort 
4 Dec 1915First published observation of filterable agent with anti-bacterial actionTwortBrown Institution
10 Sep 1917Term 'bacteriophage' coined for first timed'HerellePasteur Institute
1919First application of phage therapy to treat dysenteryd'HerelleHospital des Enfants-Malades
1919Jules Bordet, unable to replicate d'Herelle's results with phage therapy, challenged d'Herelle's conception of a phage as a virusBordet, d'Herelle 
18 Dec 1922Esther Lederberg was born in Bronx, New York, USAEsther LederbergWisconsin University
1923Establishment of Eliava Institute of Bacteriophages, Microbiology and Virology in Tbilisi, Georgiad'Herelle, EliavaGeorge Eliava Institute
1 Jan 1925Bacteriophages shown to have two different reproductive cyclesBordet, BailPasteur Institute
1925Phages successfully used to treat bubonic plagued'Herelle 
1926Modification of methods to grow phages for therapyd'HerelleHaffking Institute, India
1927Bacteriophage Inquiry launched to study the effectiveness of phage therapy 
July 1927Phage therapy tested in Kasauli, India, indicated phage therapy a promising treatment for choleraMaloneCentral Research Institute, India
August 1927Yugoslavian bacteriologist, Igor Asheshov replaced d'Herelle at the Bacteriophage Inquiryd'Herelle, Asheshov 
November 1927Phage therapy proved promising in patients with cholera in Campbell Hospital, Calcutta, Indiad'Herelle, Malone, LahiriCampbell Hospital, Kolkata, India
December 1927D'Herelle presented promising results for phage therapy to the 7th Congress of the Far Eastern Association of Tropical Medicined'Herelle 
1928 - 1929Study of phage therapy conducted in Assam, India, failed to collect sufficient evidence to prove efficacy of phage treatment for choleraMorisonKing Edward VII Pasteur Institute, India
1928Theory put forward about the contagious and hereditary nature of bacteriophagesWollmanPasteur Institute
August 1928Mixed results reported from three studies carried out with phage therapy for cholera treatmentAsheshovPuri Cholera Hospital
1929 - 1935First phage therapy programme launched in RussiaMel'nyk 
1929Igor Asheshov shifted the focus of the Bacteriophage Inquiry away from clinical application to mainly research issuesAsheshov 
8 Oct 1929Franklin W Stahl was born in Boston, Massachusetts, USAStahl California Institute of Technology, University of Missouri, University of Oregon
1931d’Herelle issued a public challenge to Bordet's theory about bacteriophagesBordet, d'HerellePasteur Institute
1931D’Herelle published his first paper on intravenous phage therapyd'HerelleYale University School of Medicine
1933Asheshov published three long articles on the classification and various properties of cholera plagues but failed to assess the effectiveness of phage therapyAsheshov 
1933Serological test established to classify phagesBurnetNational Institute for Medical Research
1933D'Herelle relocated to Georgiad'Herelle 
1934First American Medical Association (AMA) review of phage therapyEaton, BayneYale University School of Medicine
1934Eliava Institute and the Kharkov Mechnikov Institute formed a collaboration to study phages and their medical applicationsKharkov, EliavaKharkov Mechnikov Institute Eliava Institute
1935Asheshov left the Bacteriophage InquiryAsheshov 
1936Results from Bacteriophage Inquiry's last study undertaken at Campbell Hospital concluded that phage therapy 'appeared to reduce mortality by about 2.5 fold'.PasrichaCampbell Hospital, Kolkata, India
1936The Bacteriophage Inquiry ended 
July 1937George Eliava executed and d'Herelle left GeorgiaEliava, d'Herelle 
1938Soviet phage therapy used to curb outbreak of cholera in several areas of Afghanistan near the Soviet Border 
1938 - 1939The Bacteriological Institute merged with the Institute of Microbiology & Epidemiology (founded in 1936) to become the Research Institute of Microbiology, Epidemiology and BacteriophageTbilisi Institute
1939 - 1940Staphylococcal and streptococcal phages used to save Soviet soldiers wounded in the war with Finland 
1939D’Herelle imprisoned by the Germans in Paris for refusing to share his expertise on phage therapyd'Herelle 
1940First electron microscope pictures of bacteriophages publishedRuska 
1940First visualisation of a phage using electron microscopyRuskaSiemens, Halske laboratory
1941 - 1945Mass production of phage therapy launched by Soviet bacteriological institutions to treat Soviet troops wounded in World War II 
May 1941Second AMA review of phage therapyKrueger, Scribner 
22 Jun 1941Germans invaded Russia, not only to capture the oil wells but also to obtain the phages preparations at the Tbilisi Institute in GeorgiaTbilisi Institute
1942 - 1943Phage therapy proved key to the Soviet Army winning the Battle of Stalingrad 
April 1942Electron microscope used to identigy and characterise a bacteriophageLuria, Thomas AndersonColumbia University
1945Third AMA review of phage therapyMorton, Engley 
22 Feb 1949Felix d'Herelle diedd'HerellePasteur Institute
January 1950Esther Lederberg discovered the lambda phageEsther LederbergUniversity of Wisconsin
20 Mar 1950Frederick W Twort diedTwortBrown Animal Sanatory Institution
1952Ludwik Hirszfeld Institute of Immunology and Experimental Therapy establishedHirszfeldHirszfeld Institute
1963 - 1964Large-scale investigation of phage therapy for dysentery caused by Shigella undertaken by researchers at Tbilisi InstituteTbilisi Institute
1982 - 1987Phages successfully used to treat different animals infected with diarrhoea-causing E.coliHerbert Smith, HugginsHoughton Poultry Research Station
1985Phage display method is developed for selecting peptides, proteins or antibodies from a wide number of variants.SmithUniversity of Missouri
1988 - 1994Phage therapy shown to be effective in animals to control A. baumannii, P. aeruginosa, and S. aureus infections and to prevent destruction of skin grafts by P. aeruginosa Soothill, Lawrence, AyliffeUniversity of Birmingham Medical School
1989Cambridge Antibody Technology (CAT) foundedWinter, ChiswellLaboratory of Molecular Biology, CAT
1990Phage display monoclonal antibodies createdWinterLaboratory of Molecular Biology, CAT
1991First display and selection of human antibodies phageBarbas, LernerScripps Research Institute
1998Intralytix foundedSulakvelidze, MorrisIntralytix
31 Dec 2002First fully human monoclonal antibody drug approved for market WinterCAT, BASF, Abbott
2004Publication of a review on the different life cycles of bacteriophagesWeinbauerNetherlands Institute for Sea Research
August 2004First major intentional phage summit 
2006FDA approved the first bacteriophage preparation to be used as an antimicrobial agentIntralytix
11 Nov 2006Esther Lederberg diedEsther LederbergWisconsin University
2009First phase I clinical trial with phage therapy in USRhoadsSouthwest Regional Wound Care Center, Texas
2009First phase II clinical trial with phage therapy in UKWrightRoyal National Throat, Nose and Ear Hospital, London
2016FDA granted granted safe status to 'ShigaShield' phage cocktails for use in food productionIntralytix
2016Phase I clinical trial launched for phage therapy to treat S. aureus infections in patients with chronic rhinosinusitisArmata
October 2017Successful treatment of antibiotic-resistant A. baumannii infection using personalised bacteriophagesSchooley 

25 Apr 1873

Felix d'Herelle was born in Montreal, Canada

22 Oct 1877

Frederick W Twort was born in Camberley, Surrey, United Kingdom

1896

First observation of substance with potential antibacterial activity

1915

Discovery of bacteriophages, type of virus that attacks bacteria, by English bacteriologist William Twort

4 Dec 1915

First bacteriophage discovered

4 Dec 1915

First published observation of filterable agent with anti-bacterial action

10 Sep 1917

Term 'bacteriophage' coined for first time

1919

First application of phage therapy to treat dysentery

1919

Jules Bordet, unable to replicate d'Herelle's results with phage therapy, challenged d'Herelle's conception of a phage as a virus

18 Dec 1922

Esther Lederberg was born in Bronx, New York, USA

1923

Establishment of Eliava Institute of Bacteriophages, Microbiology and Virology in Tbilisi, Georgia

1 Jan 1925

Bacteriophages shown to have two different reproductive cycles

1925

Phages successfully used to treat bubonic plague

1926

Modification of methods to grow phages for therapy

1927

Bacteriophage Inquiry launched to study the effectiveness of phage therapy

Jul 1927

Phage therapy tested in Kasauli, India, indicated phage therapy a promising treatment for cholera

Aug 1927

Yugoslavian bacteriologist, Igor Asheshov replaced d'Herelle at the Bacteriophage Inquiry

Nov 1927

Phage therapy proved promising in patients with cholera in Campbell Hospital, Calcutta, India

Dec 1927

D'Herelle presented promising results for phage therapy to the 7th Congress of the Far Eastern Association of Tropical Medicine

1928 - 1929

Study of phage therapy conducted in Assam, India, failed to collect sufficient evidence to prove efficacy of phage treatment for cholera

1928

Theory put forward about the contagious and hereditary nature of bacteriophages

Aug 1928

Mixed results reported from three studies carried out with phage therapy for cholera treatment

1929 - 1935

First phage therapy programme launched in Russia

1929

Igor Asheshov shifted the focus of the Bacteriophage Inquiry away from clinical application to mainly research issues

8 Oct 1929

Franklin W Stahl was born in Boston, Massachusetts, USA

1931

d’Herelle issued a public challenge to Bordet's theory about bacteriophages

1931

D’Herelle published his first paper on intravenous phage therapy

1933

Asheshov published three long articles on the classification and various properties of cholera plagues but failed to assess the effectiveness of phage therapy

1933

Serological test established to classify phages

1933

D'Herelle relocated to Georgia

1934

First American Medical Association (AMA) review of phage therapy

1934

Eliava Institute and the Kharkov Mechnikov Institute formed a collaboration to study phages and their medical applications

1935

Asheshov left the Bacteriophage Inquiry

1936

Results from Bacteriophage Inquiry's last study undertaken at Campbell Hospital concluded that phage therapy 'appeared to reduce mortality by about 2.5 fold'.

1936

The Bacteriophage Inquiry ended

Jul 1937

George Eliava executed and d'Herelle left Georgia

1938

Soviet phage therapy used to curb outbreak of cholera in several areas of Afghanistan near the Soviet Border

1938 - 1939

The Bacteriological Institute merged with the Institute of Microbiology & Epidemiology (founded in 1936) to become the Research Institute of Microbiology, Epidemiology and Bacteriophage

1939 - 1940

Staphylococcal and streptococcal phages used to save Soviet soldiers wounded in the war with Finland

1939

D’Herelle imprisoned by the Germans in Paris for refusing to share his expertise on phage therapy

1940

First electron microscope pictures of bacteriophages published

1940

First visualisation of a phage using electron microscopy

1941 - 1945

Mass production of phage therapy launched by Soviet bacteriological institutions to treat Soviet troops wounded in World War II

May 1941

Second AMA review of phage therapy

22 Jun 1941

Germans invaded Russia, not only to capture the oil wells but also to obtain the phages preparations at the Tbilisi Institute in Georgia

1942 - 1943

Phage therapy proved key to the Soviet Army winning the Battle of Stalingrad

Apr 1942

Electron microscope used to identigy and characterise a bacteriophage

1945

Third AMA review of phage therapy

22 Feb 1949

Felix d'Herelle died

Jan 1950

Esther Lederberg discovered the lambda phage

20 Mar 1950

Frederick W Twort died

1952

Ludwik Hirszfeld Institute of Immunology and Experimental Therapy established

1963 - 1964

Large-scale investigation of phage therapy for dysentery caused by Shigella undertaken by researchers at Tbilisi Institute

1982 - 1987

Phages successfully used to treat different animals infected with diarrhoea-causing E.coli

1985

Phage display method is developed for selecting peptides, proteins or antibodies from a wide number of variants.

1988 - 1994

Phage therapy shown to be effective in animals to control A. baumannii, P. aeruginosa, and S. aureus infections and to prevent destruction of skin grafts by P. aeruginosa

1989

Cambridge Antibody Technology (CAT) founded

1990

Phage display monoclonal antibodies created

1991

First display and selection of human antibodies phage

1998

Intralytix founded

31 Dec 2002

First fully human monoclonal antibody drug approved for market

2004

Publication of a review on the different life cycles of bacteriophages

Aug 2004

First major intentional phage summit

2006

FDA approved the first bacteriophage preparation to be used as an antimicrobial agent

11 Nov 2006

Esther Lederberg died

2009

First phase I clinical trial with phage therapy in US

2009

First phase II clinical trial with phage therapy in UK

2016

FDA granted granted safe status to 'ShigaShield' phage cocktails for use in food production

2016

Phase I clinical trial launched for phage therapy to treat S. aureus infections in patients with chronic rhinosinusitis

Oct 2017

Successful treatment of antibiotic-resistant A. baumannii infection using personalised bacteriophages

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