Generation, Diversity Determination, and Application to Antibody Selection of a Human Naïve Fab Library

Kim, Sangkyu; Park, Insoo; Park, Seung Gu; Cho, Seulki; Kim, Jin Hong; Ipper, Nagesh S.; Choi, Sun Shim; Lee, Eung Suk; Hong, Hyo Jeong

doi:10.14348/molcells.2017.0106

Cited by 21 publications

(14 citation statements)

References 52 publications

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“…These libraries are generated from healthy donor's mRNA or RNA without bias toward a particular disease state, and are used to yield mAbs against unlimited range of antigens ( 151 ). To generate a highly diverse naïve antibody phage library, it is recommended to use a large pool of donors from diverse ethnic groups, and to maximize the efficiency of antibody gene amplification in the process of library construction ( 152 – 155 ).…”

Section: Types Of Antibody Phage Display Librariesmentioning

confidence: 99%

Phage Display Derived Monoclonal Antibodies: From Bench to Bedside

et al. 2020

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Monoclonal antibodies (mAbs) have become one of the most important classes of biopharmaceutical products, and they continue to dominate the universe of biopharmaceutical markets in terms of approval and sales. They are the most profitable single product class, where they represent six of the top ten selling drugs. At the beginning of the 1990s, an in vitro antibody selection technology known as antibody phage display was developed by John McCafferty and Sir. Gregory Winter that enabled the discovery of human antibodies for diverse applications, particularly antibody-based drugs. They created combinatorial antibody libraries on filamentous phage to be utilized for generating antigen specific antibodies in a matter of weeks. Since then, more than 70 phage–derived antibodies entered clinical studies and 14 of them have been approved. These antibodies are indicated for cancer, and non-cancer medical conditions, such as inflammatory, optical, infectious, or immunological diseases. This review will illustrate the utility of phage display as a powerful platform for therapeutic antibodies discovery and describe in detail all the approved mAbs derived from phage display.

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Section: Types Of Antibody Phage Display Librariesmentioning

confidence: 99%

Phage Display Derived Monoclonal Antibodies: From Bench to Bedside

et al. 2020

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“…The percentage expression (or display) was calculated based on the number of clones with an OD (optical density) over three-fold the background in an ELISA (Enzyme-Linked ImmunoSorbent Assay) for periplasmic expression of Fab or scFv. Kim et al [28] reported as functional ORFs those sequences with no stop codons; they did not report the expression nor display before the selections. The authors of pIX V3.0 [29] estimated the percentage of ORFs as sequences without stop codons and functionality as Fab display in an ELISA.…”

Section: Effective Size Of the Phage Display Antibody Librariesmentioning

confidence: 99%

“…It combines kappa (HAL10) and lambda (HAL9) light chains displayed as scFvs. The fourth and most recently published naïve library [28] was developed at Kangwon National University (KNU). It only contains kappa-type light chains, and thus we called it KNU-Fab.…”

Section: Types Of Antibody Repertoiresmentioning

confidence: 99%

Phage Display Libraries for Antibody Therapeutic Discovery and Development

et al. 2019

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Phage display technology has played a key role in the remarkable progress of discovering and optimizing antibodies for diverse applications, particularly antibody-based drugs. This technology was initially developed by George Smith in the mid-1980s and applied by John McCafferty and Gregory Winter to antibody engineering at the beginning of 1990s. Here, we compare nine phage display antibody libraries published in the last decade, which represent the state of the art in the discovery and development of therapeutic antibodies using phage display. We first discuss the quality of the libraries and the diverse types of antibody repertoires used as substrates to build the libraries, i.e., naïve, synthetic, and semisynthetic. Second, we review the performance of the libraries in terms of the number of positive clones per panning, hit rate, affinity, and developability of the selected antibodies. Finally, we highlight current opportunities and challenges pertaining to phage display platforms and related display technologies.

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“…For natural naive libraries, the general paradigm has been to increase the natural source of B-cell diversity by increasing the number of donors 23 , 25 , 26 . The question remains as to whether increasing the number of donors actually leads to improved antibody diversity and library functionality.…”

Section: Introductionmentioning

confidence: 99%

A single donor is sufficient to produce a highly functional in vitro antibody library

et al. 2021

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Antibody complementarity determining region diversity has been considered to be the most important metric for the production of a functional antibody library. Generally, the greater the antibody library diversity, the greater the probability of selecting a diverse array of high affinity leads. According to this paradigm, the primary means of elevating library diversity has been by increasing the number of donors. In the present study we explored the possibility of creating an in vitro antibody library from a single healthy individual, showing that the number of lymphocytes, rather than the number of donors, is the key criterion in the production of a diverse and functional antibody library. We describe the construction of a high-quality phage display library comprising 5 × 109 human antibodies by applying an efficient B cell extraction protocol from a single donor and a targeted V-gene amplification strategy favoring specific antibody families for their improved developability profiles. Each step of the library generation process was followed and validated by next generation sequencing to monitor the library quality and diversity. The functionality of the library was tested using several therapeutically relevant targets for which a vast number of different antibodies with desired biophysical properties were obtained.

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Generation, Diversity Determination, and Application to Antibody Selection of a Human Naïve Fab Library

Cited by 21 publications

References 52 publications

Phage Display Derived Monoclonal Antibodies: From Bench to Bedside

Phage Display Derived Monoclonal Antibodies: From Bench to Bedside

Phage Display Libraries for Antibody Therapeutic Discovery and Development

A single donor is sufficient to produce a highly functional in vitro antibody library

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