Phage-display technology for the production of recombinant monoclonal antibodies

Rajput, Roopali; Khanna, Madhu; Pradhan, H.K.

doi:10.13070/mm.en.4.873

Cited by 5 publications

(5 citation statements)

References 109 publications

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“…The occurrence of frequent flu outbreaks leads to large-scale socio-economic losses and calls for improvement in the production of reagents, in terms of large scale production, uniform quality, low cost, and stability for effective diagnosis and management of the disease. With the advent of upgraded technologies, rAbs have provided great advantages against the conventional monoclonal antibodies ( 28 , 29 ). Once developed, rAbs are economically one of the most feasible diagnostic and therapeutic agents and allow mass scale production.…”

Section: Discussionmentioning

confidence: 99%

Diagnostic Potential of Recombinant scFv Antibodies Generated Against Hemagglutinin Protein of Influenza A Virus

et al. 2015

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Human influenza A viruses have been the cause of enormous socio-economic losses worldwide. In order to combat such a notorious pathogen, hemagglutinin protein (HA) has been a preferred target for generation of neutralizing-antibodies as potent therapeutic/diagnostic agents. In the present study, recombinant anti-HA single chain variable fragment antibodies were constructed using the phage-display technology to aid in diagnosis and treatment of human influenza A virus infections. Spleen cells of mice hyper-immunized with A/New Caledonia/20/99 (H1N1) virus were used as the source for recombinant antibody (rAb) production. The antigen-binding phages were quantified after six rounds of bio-panning against A/New Caledonia/20/99 (H1N1), A/California/07/2009 (H1N1)-like, or A/Udorn/307/72(H3N2) viruses. The maximum phage yield was for the A/New Caledonia/20/99 (H1N1), however, considerable cross-reactivity was observed for the other virus strains as well. The HA-specific polyclonal rAb preparation was subjected to selection of single clones for identification of high reactive relatively conserved epitopes. The high-affinity rAbs were tested against certain known conserved HA epitopes by peptide ELISA. Three recombinant mAbs showed reactivity with both the H1N1 strains and one (C5) showed binding with all the three viral strains. The C5 antibody was thus used for development of an ELISA test for diagnosis of influenza virus infection. Based on the sample size in the current analysis, the ELISA test demonstrated 83.9% sensitivity and 100% specificity. Thus, the ELISA, developed in our study, may prove as a cheaper alternative to the presently used real time RT–PCR test for detection of human influenza A viruses in clinical specimens, which will be beneficial, especially in the developing countries.

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Section: Discussionmentioning

confidence: 99%

Diagnostic Potential of Recombinant scFv Antibodies Generated Against Hemagglutinin Protein of Influenza A Virus

et al. 2015

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“…The diversity of antibodies from antibody repertoires is dependent on the initially estimated complexity of the fragment gene sequence library. Since the antigen-binding region is a heterodimer, the complexity of the library is enriched by the random fusion of heavy-chain and light-chain sub-libraries (40).…”

Section: Display Technologies In Mab Productionmentioning

confidence: 99%

“…This is one of the most essential features of a recombinant DNA vector. A phagemid vector provides these essential features and allows the expression of foreign DNA which is intentionally inserted in the vector as expressed in fusion with a phage protein to be displayed on the surface of the phage (40). A variety of bacteriophages have been used in phage display, most commonly used were filamentous bacteriophages which are highly productive.…”

Section: Display Technologies In Mab Productionmentioning

confidence: 99%

A Review on The Development, Production Strategies, and Utilization of Monoclonal Antibodies

Şanlav¹,

Bekçioğlu²,

Başbınar³

2020

J Basic Clin Health Sci

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Over the last 20 years, monoclonal antibodies have become the backbone of biological therapeutics for the treatment and diagnosis of several diseases. The rising incidence of cancer and other immunologic diseases promoted the increasing investments of the global pharmaceutical industry in monoclonal antibody development. The R&D has focused on the highest efficacy which is majorly correlated with the antigenbinding specificity and the lowest immunogenicity of monoclonal antibodies. This review aims to provide a brief description and explanation of each stage in the development path of mAbs.

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“… 3 , 4 The difficulty in controlling stringency during binding causes many candidates from the enriched phage pools to represent false positives that fail in vitro validation. 5 , 6 An inability to recapitulate the low relative concentration and morphology of targets in vitro may also lead to candidates failing later during in vivo testing. 7 The stochastic nature of selection results in thousands of nonspecific clones, requiring further screening for elimination.…”

Section: Introductionmentioning

confidence: 99%

“…Conventionally, >5 rounds of selection are required to generate clones with high affinity to the target. The cost and time requirements of repeat rounds (approximately 6–8 weeks and $8,000–10,000 USD in total) are a bottleneck in the discovery of new therapeutics. , The difficulty in controlling stringency during binding causes many candidates from the enriched phage pools to represent false positives that fail in vitro validation. , An inability to recapitulate the low relative concentration and morphology of targets in vitro may also lead to candidates failing later during in vivo testing . The stochastic nature of selection results in thousands of nonspecific clones, requiring further screening for elimination.…”

Section: Introductionmentioning

confidence: 99%

Rapid On-Cell Selection of High-Performance Human Antibodies

et al. 2021

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Phage display is a critical tool for developing antibodies. However, existing approaches require many time-consuming rounds of biopanning and screening of potential candidates due to a high rate of failure during validation. Herein, we present a rapid on-cell phage display platform which recapitulates the complex in vivo binding environment to produce high-performance human antibodies in a short amount of time. Selection is performed in a highly stringent heterogeneous mixture of cells to quickly remove nonspecific binders. A microfluidic platform then separates antigen-presenting cells with high throughput and specificity. An unsupervised machine learning algorithm analyzes sequences of phage from all pools to identify the structural trends that contribute to affinity and proposes ideal candidates for validation. In a proof-of-concept screen against human Frizzled-7, a key ligand in the Wnt signaling pathway, antibodies with picomolar affinity were discovered in two rounds of selection that outperformed current gold-standard reagents. This approach, termed μCellect, is low cost, high throughput, and compatible with a wide variety of cell types, enabling widespread adoption for antibody development.

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Phage-display technology for the production of recombinant monoclonal antibodies

Cited by 5 publications

References 109 publications

Diagnostic Potential of Recombinant scFv Antibodies Generated Against Hemagglutinin Protein of Influenza A Virus

Diagnostic Potential of Recombinant scFv Antibodies Generated Against Hemagglutinin Protein of Influenza A Virus

A Review on The Development, Production Strategies, and Utilization of Monoclonal Antibodies

Rapid On-Cell Selection of High-Performance Human Antibodies

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