Llama antibody fragments with cross-subtype human immunodeficiency virus type 1 (HIV-1)-neutralizing properties and high affinity for HIV-1 gp120

Anna Forsman, Els Beirnaert, Marlén M. I. Aasa-Chapman, Bart Hoorelbeke, Karolin Hijazi, Willie Koh, Vanessa Tack, Agnieszka Szynol, Charles Kelly, Aine McKnight, Theo Verrips, Hans de Haard, Robin A Weiss

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Abstract

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.
Original languageEnglish
Pages (from-to)12069-12081
Number of pages13
JournalJournal of Virology
Volume82
Issue number24
Early online date8 Oct 2008
DOIs
Publication statusPublished - Dec 2008

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New World Camelids
Immunoglobulin Fragments
llamas
Human immunodeficiency virus 1
neutralization
HIV-1
antibodies
Human Immunodeficiency Virus env Gene Products
Camelidae
inhibitory concentration 50
immunoglobulin light chains
Immunoglobulin Light Chains
Virus Internalization
compound A 12
Antibodies
DNA libraries
Virus Diseases
Anti-Infective Agents
Bacteriophages
epitopes

Keywords

  • animals
  • antibodies
  • antigens, CD4
  • binding sites
  • camelids, new world
  • cross reactions
  • epitopes
  • HIV envelope protein gp120
  • HIV-1
  • humans
  • recombinant proteins

Cite this

Llama antibody fragments with cross-subtype human immunodeficiency virus type 1 (HIV-1)-neutralizing properties and high affinity for HIV-1 gp120. / Forsman, Anna; Beirnaert, Els; Aasa-Chapman, Marlén M. I.; Hoorelbeke, Bart; Hijazi, Karolin; Koh, Willie; Tack, Vanessa; Szynol, Agnieszka; Kelly, Charles; McKnight, Aine; Verrips, Theo; de Haard, Hans; Weiss, Robin A.

In: Journal of Virology, Vol. 82, No. 24, 12.2008, p. 12069-12081.

Research output: Contribution to journalArticle

Forsman, A, Beirnaert, E, Aasa-Chapman, MMI, Hoorelbeke, B, Hijazi, K, Koh, W, Tack, V, Szynol, A, Kelly, C, McKnight, A, Verrips, T, de Haard, H & Weiss, RA 2008, 'Llama antibody fragments with cross-subtype human immunodeficiency virus type 1 (HIV-1)-neutralizing properties and high affinity for HIV-1 gp120', Journal of Virology, vol. 82, no. 24, pp. 12069-12081. https://doi.org/10.1128/JVI.01379-08
Forsman, Anna ; Beirnaert, Els ; Aasa-Chapman, Marlén M. I. ; Hoorelbeke, Bart ; Hijazi, Karolin ; Koh, Willie ; Tack, Vanessa ; Szynol, Agnieszka ; Kelly, Charles ; McKnight, Aine ; Verrips, Theo ; de Haard, Hans ; Weiss, Robin A. / Llama antibody fragments with cross-subtype human immunodeficiency virus type 1 (HIV-1)-neutralizing properties and high affinity for HIV-1 gp120. In: Journal of Virology. 2008 ; Vol. 82, No. 24. pp. 12069-12081.
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abstract = "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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T1 - Llama antibody fragments with cross-subtype human immunodeficiency virus type 1 (HIV-1)-neutralizing properties and high affinity for HIV-1 gp120

AU - Forsman, Anna

AU - Beirnaert, Els

AU - Aasa-Chapman, Marlén M. I.

AU - Hoorelbeke, Bart

AU - Hijazi, Karolin

AU - Koh, Willie

AU - Tack, Vanessa

AU - Szynol, Agnieszka

AU - Kelly, Charles

AU - McKnight, Aine

AU - Verrips, Theo

AU - de Haard, Hans

AU - Weiss, Robin A

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N2 - 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.

AB - 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.

KW - animals

KW - antibodies

KW - antigens, CD4

KW - binding sites

KW - camelids, new world

KW - cross reactions

KW - epitopes

KW - HIV envelope protein gp120

KW - HIV-1

KW - humans

KW - recombinant proteins

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DO - 10.1128/JVI.01379-08

M3 - Article

C2 - 18842738

VL - 82

SP - 12069

EP - 12081

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 24

ER -