Isolation of Monoclonal Antiobodies Against HIV

National Institute Of Allergy And Infectious Diseases

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Abstract

Some HIV infected patients develop potent antibody responses against HIV, but it is not clear how often this happens, nor how it occurs. We have undertaken a systemic evaluation of HIV+ sera to find those with broad neutralizing activity, understand how breadth of neutralization occurs and to isolate antibodies from the cells of such patients. [unreadable] [unreadable] Over the last year we have screened over 100 patient sera and found that about 20% have broadly neutralizing antibodies. Among this group, we selected those patients with the broadest antibody response and have collected PBMC in order to isolate monoclonal antibodies. We have takes several approaches to achieve this, including collaborations with Dr. Antonio Lanzevecchia and Michel Nussenzweig. The Lanzevecchia approach involves isolation of memory B-cells, EBV transformation and screening for HIV specific cells. The Nussenzweig approach involves isolating memory B-cells that are HIV-1 specific, and performing single cell PCR to isolate IgG genes. The genes are then cloned and expressed as full IgGs. To date, these approaches have yielded several novel anti-HIV monoclonal antibodies that we are in the process of evaluating.