Document 0166
 DOCN  M94A0166
 TI    Generation of human monoclonal antibodies against HIV-1 proteins;
       electrofusion and Epstein-Barr virus transformation for peripheral blood
       lymphocyte immortalization.
 DT    9412
 AU    Buchacher A; Predl R; Strutzenberger K; Steinfellner W; Trkola A;
       Purtscher M; Gruber G; Tauer C; Steindl F; Jungbauer A; et al; Institute
       of Applied Microbiology, University of Foresty and; Agriculture, Vienna,
       Austria.
 SO    AIDS Res Hum Retroviruses. 1994 Apr;10(4):359-69. Unique Identifier :
       AIDSLINE MED/94347460
 AB    Electrofusion and EBV transformation were studied by immortalizing human
       PBLs from blood of HIV-1-positive volunteers. A panel of 33 cell lines
       producing human monoclonal antibodies (Hu-MAbs) against HIV-1 was
       established by cell fusion or EBV transformation. For the first fusion
       experiments the source of B lymphocytes was peripheral blood of
       HIV-1-infected donors in CDC stages II or III with CD4 cell counts
       higher than 500/mm3. Later on, from these patients only, those with high
       anti-HIV titers were chosen as blood donors. By that means the yield of
       stable specific hybridomas was increased twofold. In our experiments
       electrofusion turned out to be a more efficient immortalization method
       than EBV transformation, due to a high and constant immortalization
       rate. The hybridomas were stable after intensive subcloning and could be
       cultivated over a period of 8 months without loss in monoclonal antibody
       production. Immunoglobulin class, subtype, reactivity against HIV-1
       proteins, Western blot patterns, immunofluorescence, and epitopes were
       characterized. The subtype of all antibodies was IgG1 or IgG3. The light
       chain was predominantly kappa. All antibodies showed reactivity against
       HIV-1 envelope or core protein. All hybridomas were stable and suited
       for mass production. Several Hu-MAbs are becoming an important tool in
       the field of diagnosis, research, and immunotherapy.
 DE    Antibodies, Monoclonal/*BIOSYNTHESIS/CHEMISTRY  Antigenic Determinants
       B-Lymphocytes/IMMUNOLOGY  Cell Fusion  Cell Line  Cell Transformation,
       Viral  Herpesvirus 4, Human  Human  Hybridomas/IMMUNOLOGY  HIV
       Antibodies/*BIOSYNTHESIS/CHEMISTRY  HIV Envelope Protein
       gp120/IMMUNOLOGY  HIV Infections/IMMUNOLOGY  HIV-1/*IMMUNOLOGY
       Immunochemistry  In Vitro  Neutralization Tests  JOURNAL ARTICLE

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