Generation of Human Monoclonal Antibodies against HIV-1 Proteins; Electrofusion and Epstein-Barr Virus Transformation for Peripheral Blood Lymphocyte Immortalization

Buchacher, Andrea; Predl, R.; Strutzenberger, K.; Steinfellner, W.; Trkola, Alexandra; Purtscher, Martin; Grüber, Gerhard; Tauer, Christa; Steindl, Franz; Jungbauer, Alois; Katinger, Hermann

doi:10.1089/aid.1994.10.359

Cited by 505 publications

(366 citation statements)

References 32 publications

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“…Of these, only a handful have been found to be broadly neutralizing across HIV-1 subtypes (8), and have been the result of HIV-1 infection rather than immunization. Two of these are directed against gp120: MAb b12, which binds to an epitope that overlaps a subset of the CD4-binding site (CD4bs) of gp120 (3,10,11,68,94), and MAb 2G12, which recognizes a carbohydrate motif (9,70,72,80). Two broadly neutralizing MAbs, 4E10 and 2F5, recognize gp41 (9,56,77,96).…”

mentioning

confidence: 99%

“…Two of these are directed against gp120: MAb b12, which binds to an epitope that overlaps a subset of the CD4-binding site (CD4bs) of gp120 (3,10,11,68,94), and MAb 2G12, which recognizes a carbohydrate motif (9,70,72,80). Two broadly neutralizing MAbs, 4E10 and 2F5, recognize gp41 (9,56,77,96). MAbs X5 (55), which recognizes an epitope on gp120 that is better-exposed after CD4 binding, and m14 (92), which competes with CD4 for binding to gp120, also display some neutralizing activity across HIV-1 subtypes, as do a minority of MAbs to the V3 region of gp120 (30,31).…”

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confidence: 99%

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Llama Antibody Fragments with Cross-Subtype Human Immunodeficiency Virus Type 1 (HIV-1)-Neutralizing Properties and High Affinity for HIV-1 gp120

Forsman

Beirnaert

Aasa-Chapman

et al. 2008

J Virol

111

138

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Members of the Camelidae family produce immunoglobulins devoid of light chains. We have characterized variable domains of these heavy chain antibodies, the VHH, from llamas immunized with human immunodeficiency virus type 1 (HIV-1) envelope protein gp120 in order to identify VHH that can inhibit HIV-1 infection. To increase the chances of isolating neutralizing VHH, we employed a functional selection approach, involving panning of phage libraries expressing the VHH repertoire on recombinant gp120, followed by a competitive elution with soluble CD4. By immunizing with gp120 derived from an HIV-1 subtype B /C primary isolate, followed by panning on gp120 from HIV-1 isolates of subtypes A, B, and C, we could select for VHH with cross-subtype neutralizing activity. Three VHH able to neutralize HIV-1 primary isolates of subtypes B and C were characterized. These bound to recombinant gp120 with affinities close to the suggested affinity ceiling for in vivo-maturated antibodies and competed with soluble CD4 for this binding, indicating that their mechanism of neutralization involves interacting with the functional envelope spike prior to binding to CD4. The most potent VHH in terms of low 50% inhibitory concentration (IC 50 ) and IC 90 values and cross-subtype reactivity was A12. These results indicate that camelid VHH can be potent HIV-1 entry inhibitors. Since VHH are stable and can be produced at a relatively low cost, they may be considered for applications such as HIV-1 microbicide development. Antienvelope VHH might also prove useful in defining neutralizing and nonneutralizing epitopes on HIV-1 envelope proteins, with implications for HIV-1 vaccine design.

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confidence: 99%

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confidence: 99%

Llama Antibody Fragments with Cross-Subtype Human Immunodeficiency Virus Type 1 (HIV-1)-Neutralizing Properties and High Affinity for HIV-1 gp120

Forsman

Beirnaert

Aasa-Chapman

et al. 2008

J Virol

111

138

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“…These CDRH3 regions help the antibodies to penetrate the glycan shield of Env trimer then interact with the V1/V2 and/or V3 region of gp120 [73]. Also, CDRH3 is an important component in gp41-reactive antibodies such as 2F5 (CDRH3 of 22 residues) and 4E10 (CDRH3 of 18 residues) which performs the additional activity causing a hydrophobic interaction with the membrane [74][75][76][77][78][79][80] and forming a loop to contact the highly conserved hydrophobic residues on gp41 [52,[81][82][83]. …”

Section: Bovine Cdrh3 Length Goes Beyond Expectationmentioning

confidence: 99%

Broad Neutralizing Antibodies to HIV Env and Other Complex Viral Antigens from Vaccinated Cows

Heydarchi¹,

Quiroz²,

Purcell³

2016

J Vaccines Vaccin

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The idea of using antibodies to restrain HIV viral replication has been a growing interest since many years ago. HIV-patient serum with elite virus-neutralizing breadth has led to the preparation of broadly neutralizing antibodies (BrNAbs) with long and highly mutated CDRH3 domains that can neutralize a broad array of viral strains and prevent transmission in animal models. Recent advances have resulted in the discovery of BrNAbs that are more potent and can neutralize many HIV-1 subtypes. However, elicitation of these antibodies in infected individuals usually requires a long time of antigen exposure. Though BrNAbs are shown to be successful either therapeutically or prophylactically against HIV-1, production of these antibodies in bulk, commercial-sized batches are expensive and non-affordable particularly for poor countries. Therefore, more durable preventative or therapeutic strategies are required. Immunization of the cows with HIV Env is shown to be capable of producing 20 kg purified anti-HIV-1 BrNAbs and this amount could be sufficient for 2 million × 10 mg doses for formulation and pre-clinical testing as an HIV microbicide. In addition, bovine immunoglobulins typically have variable third heavy complementarity determining regions (CDRH3) that may potentially facilitate access to antigenic epitopes that are very difficult for other species to engage. Thus, cows could be engaged to elicit anti-HIV antibodies with the features of human BrNAbs and bovine colostrum could be a promising and cheap resource for the development of combination microbicides.

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“…Recombinant HIV-1 gp160 and gp41 glycoproteins (both HIV-1, MN) were from Protein Sciences Corporation (Meriden, CT). The following reagents were obtained through the AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH: Human MAb to HIV (Md-1) from Robert A. Meyers; HIV-1 gp41 MAb (2F5) from Dr. Hermann Katinger [28][29][30] ; and Chessie 8 from Dr. George Lewis. 31 …”

Section: Virus Cell Lines Glycoproteins and Mabsmentioning

confidence: 99%

Quaternary protein mimetics of gp41 elicit neutralizing antibodies against HIV fusion‐active intermediate state

Sadler

Zhang

et al. 2008

Biopolymers

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During viral entry, the fusogenic state of human immunodeficiency virus Type 1 (HIV‐1) envelope protein gp41 is a quaternary structure consisting of three gp41 glycoproteins, each with two conserved helical domains (N‐HR and C‐HR). Thus far, the examination of monomeric gp41 peptides as an immunologically focused approach to vaccine design has not been successful. Here we report an approach using quaternary protein mimetics (called 3α mimetics) that are based on the gp41 N‐HR and C‐HR domains to closely mimic the fusogenic state and overcome the deficiencies of the monomeric peptide approach for synthetic vaccine design. The 3α mimetics are conveniently prepared by chemoselective ligation of unprotected monomeric peptides to an interstrand linker, and display enhanced conformational stability compared to the corresponding monomers. The 3α mimetics with or without a covalently attached T‐helper epitope were immunogenic and elicited antisera that bound both recombinant gp160, which contains gp41, and HIV‐1 virions and immunoprecipitated recombinant gp41. Anti‐3α mimetic antisera neutralized viral infectivity against R5‐ and X4‐tropic strains of HIV‐1 at 31.5°C. The results suggest that a quaternary protein approach to mimic conserved and functional domains of viral envelope proteins is desirable for HIV vaccine development as such antigens are more likely to produce immunologically‐focused and broadly neutralizing antibody responses. © 2008 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 90: 320–329, 2008.This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com

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Generation of Human Monoclonal Antibodies against HIV-1 Proteins; Electrofusion and Epstein-Barr Virus Transformation for Peripheral Blood Lymphocyte Immortalization

Cited by 505 publications

References 32 publications

Llama Antibody Fragments with Cross-Subtype Human Immunodeficiency Virus Type 1 (HIV-1)-Neutralizing Properties and High Affinity for HIV-1 gp120

Llama Antibody Fragments with Cross-Subtype Human Immunodeficiency Virus Type 1 (HIV-1)-Neutralizing Properties and High Affinity for HIV-1 gp120

Broad Neutralizing Antibodies to HIV Env and Other Complex Viral Antigens from Vaccinated Cows

Quaternary protein mimetics of gp41 elicit neutralizing antibodies against HIV fusion‐active intermediate state

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