Document 0253
 DOCN  M9460253
 TI    Studies with monoclonal antibodies to the V3 region of HIV-1 gp120
       reveal limitations to the utility of solid-phase peptide binding assays.
 DT    9404
 AU    Moore JP; Cao Y; Conley AJ; Wyatt R; Robinson J; Gorny MK; Zolla-Pazner
       S; Ho DD; Koup RA; Aaron Diamond AIDS Research Center, New York
       University School of; Medicine, NY 10016.
 SO    J Acquir Immune Defic Syndr. 1994 Apr;7(4):332-9. Unique Identifier :
       AIDSLINE MED/94180306
 AB    Using human monoclonal antibodies (HuMAbs) r(1)-447 (L-736,523) and 19b
       to the V3 region of HIV-1 gp120, we have explored epitope presentation
       on V3-peptides and on the corresponding gp120 proteins. HuMAb r(1)-447
       binds strongly to the MN and SF-2 peptides and gp120 proteins. In
       contrast, while this HuMAb binds equally avidly to both the HxB2 and the
       BRU/BH10 peptides, it binds but weakly to the HxB2 V3 loop on gp120 and
       fails to bind at all to BH10 gp120. Thus, the solid-phase peptide
       binding assay can falsely predict reactivity of an MAb with a gp120
       protein. Conversely, HuMAb 19b fails to bind to a peptide from the V3
       loop of HIV-1 AD-6 in solid-phase assays, but binds to the same peptide
       in solution and also to AD-6 gp120. Thus, the solid-phase peptide
       binding assay can fail to predict reactivity of an MAb with a gp120
       protein. Furthermore, serum antibodies from individual AD-6 do not react
       well with the AD-6 V3-peptide in a solid-phase assay, but react strongly
       with the corresponding MN V3-peptide. On the basis of peptide binding
       assays, we had assumed that the AD-6 virus was MN-like with a prototypic
       North American/European subtype B GPGR motif at the crown of the V3
       loop. However, direct sequencing demonstrates that the AD-6 V3 loop
       contains a variant GPGK motif. This highlights a limitation of
       V3-peptide-based assays for serotyping viruses.
 DE    Amino Acid Sequence  Antibodies, Monoclonal/*IMMUNOLOGY  Antigenic
       Determinants/IMMUNOLOGY  Binding, Competitive  Enzyme-Linked
       Immunosorbent Assay  Human  HIV Envelope Protein
       gp120/CHEMISTRY/*IMMUNOLOGY  HIV-1/CLASSIFICATION/*IMMUNOLOGY  Molecular
       Sequence Data  Peptide Fragments/CHEMISTRY/*IMMUNOLOGY  Polymorphism
       (Genetics)  Recombinant Proteins/IMMUNOLOGY  Sensitivity and Specificity
       Serotyping  Support, Non-U.S. Gov't  Support, U.S. Gov't, Non-P.H.S.
       Support, U.S. Gov't, P.H.S.  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).