Isolation of Monoclonal Antiobodies Against HIV

一些HIV感染者会产生针对HIV的强有力的抗体反应，但目前尚不清楚这种情况发生的频率，也不清楚它是如何发生的。我们对HIV+血清进行了系统性评价，以发现具有广泛中和活性的血清，了解中和的广度如何发生，并从此类患者的细胞中分离抗体。 在过去的一年里，我们筛选了100多名患者的血清，发现约20%的血清具有广泛的中和抗体。在该组中，我们选择了具有最广泛抗体应答的患者，并收集PBMC以分离单克隆抗体。我们的方法包括分离HIV-1特异性的记忆B细胞，并进行单细胞PCR以分离IgG基因。然后将基因克隆并表达为完整IgG。到目前为止，这些方法已经产生了几种新的抗HIV单克隆抗体，我们正在评估。

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. 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. Our methodology includes 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.