A Novel Approach to Human Anti-HLA mABs Production: Use of Phage Display Libraries

Pistillo, Maria Pia; Hammer, Juergen; Bono, Elisa; Sinigaglia, Francesco; Bottero, Fabio; Gho, Andrea; Ferrara, Giovanni Battista

doi:10.1016/s0198-8859(97)00176-6

Cited by 7 publications

(2 citation statements)

References 29 publications

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“…One major problem of using purified antigens to isolate cell surface antibodies is that the conformation of many membrane proteins is modified by purification. Selection may isolate antibodies reactive with the purified antigen that do not react with membrane–bound antigen in its normal stoichiometry [43]. …”

Section: Selection Of Cell–surface–reactive Antibodies From Phage–dismentioning

confidence: 99%

Introduction to Antibody Engineering and Phage Display

Watkins

Ouwehand

2000

Vox Sanguinis

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The vertebrate immune system, capable of rapidly producing highly specific antibodies upon immunisation, has been used to produce murine monoclonal antibodies (mAbs) via immortalisation and isolation of antibody–secreting cells. These mAbs have had a profound impact in the fields of diagnostics and therapeutics. However, the therapeutic use of murine mAbs is complicated by their immunogenicity. To circumvent this immunogenicity, antibody engineering techniques which render murine mAbs more compatible with the human immune system have been devised. Over the last decade, the technique of antibody phage display, which facilitates the production of human mAbs, has been developed. This review serves as an introduction to antibody engineering, phage display and the development of antibody fragments into viable diagnostic and therapeutic reagents.

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Section: Selection Of Cell–surface–reactive Antibodies From Phage–dismentioning

confidence: 99%

Introduction to Antibody Engineering and Phage Display

Watkins

Ouwehand

2000

Vox Sanguinis

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“…Advances in immortalizing human B cells and the development of phage-display technologies allowed for the isolation of anti-HLA monocolonal antibodies from allosensitized individuals [10, 11]. The anti-HLA antibodies have proven to be valuable reagents for investigating MHC molecules themselves and their role in disease processes.…”

Section: Introductionmentioning

confidence: 99%

Isolation of a monoclonal antibody from a phage display library binding the rhesus macaque MHC class I allomorph Mamu-A1*001

et al. 2017

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Monoclonal antibodies that bind to human leukocyte antigen (HLA) are useful tools for HLA-typing, tracking donor-recipient chimerisms after bone marrow transplants, and characterizing specific major histocompatibility complexes (MHC) on cell surfaces. Unfortunately, equivalent reagents are not available for rhesus macaques, which are commonly used animal as models in organ transplant and infectious disease research. To address this deficiency, we isolated an antibody that recognizes the common Indian rhesus macaque MHC class I molecule, Mamu-A1*001. We induced Mamu-A1*001-binding antibodies by alloimmunizing a female Mamu-A1*001-negative rhesus macaque with peripheral blood mononuclear cells (PBMC) from a male Mamu-A1*001-positive donor. A Fab phage display library was constructed with PBMC from the alloimmunized macaque and panned to isolate an antibody that binds to Mamu-A1*001 but not to other common rhesus macaque MHC class I molecules. The isolated antibody distinguishes PBMC from Mamu-A1*001-positive and -negative macaques. Additionally, the Mamu-A1*001-specific antibody binds the cynomolgus macaque MHC class I ortholog Mafa-A1*001:01 but not variants Mafa-A1*001:02/03, indicating a high degree of binding specificity. The Mamu-A1*001-specific antibody will be useful for identifying Mamu-A1*001-positive rhesus macaques, for detecting Mamu-A1*001-positive cells in populations of Mamu-A1*001-negative cells, and for examining disease processes that alter expression of Mamu-A1*001 on cell surfaces. Moreover, the alloimmunization process we describe will be useful for isolating additional MHC allomorph-specific monoclonal antibodies or antibodies against other polymorphic host proteins which are difficult to isolate with traditional technologies.

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