A critical aspect of this program project includes evaluating antibodies for their potency against HIV and affinities to gpl 20s and Fc receptors. The Cell/Biochemical Assay Automation Core will design and carry out cell-based and biochemical assays to characterize the specificity, efficacy, and potency of HIV-neutralizing antibodies. We have adapted the standard in vitro pseudovirus neutralization assay, which measures the reduction in luciferase reporter gene expression in the presence of a potential inhibitor following pseudovirus infection in TZM-bl cells, for execution by a liquid handing robot. Core A personnel will validate, refine, and trouble-shoot protocols for the automated in vitro neutralization assays and then perform automated in vitro neutralization assays for evaluating antibodies for Drs. Nussenzweig, Ravetch, and Bjorkman. In addition. Core A will perform rapid and high-throughput binding assays to assess the affinities of antibodies for gpl20s and/or gp140s, as required for Drs. Nussenzweig and Bjorkman, and the affinities of modified Fc regions for Fc receptors, as required by Dr. Ravetch. The core will train and assist students, postdoctoral fellows, research assistants, and investigators in the analysis and interpretation of data from automated assays. In addition, the core will maintain two custom-equipped Evo Freedom Liquid handling stations, including daily calibration and care, and annual preventative maintenance.
HIV/AIDS is a global public health problem. Efforts to develop a vaccine or improve antibodies for passive immunization require screening large numbers of potential reagents: using in vitro neutralization assays to evaluate antibody potencies and protein-protein binding assays to identify interactions. We have automated these techniques in order to efficiently evaluate large numbers of reagents for the investigators in this collaborative project.
| Wang, Haoqing; Barnes, Christopher O; Yang, Zhi et al. (2018) Partially Open HIV-1 Envelope Structures Exhibit Conformational Changes Relevant for Coreceptor Binding and Fusion. Cell Host Microbe 24:579-592.e4 |
| Stadtmueller, Beth M; Bridges, Michael D; Dam, Kim-Marie et al. (2018) DEER Spectroscopy Measurements Reveal Multiple Conformations of HIV-1 SOSIP Envelopes that Show Similarities with Envelopes on Native Virions. Immunity 49:235-246.e4 |
| Gautam, Rajeev; Nishimura, Yoshiaki; Gaughan, Natalie et al. (2018) A single injection of crystallizable fragment domain-modified antibodies elicits durable protection from SHIV infection. Nat Med 24:610-616 |
| Cohn, Lillian B; da Silva, Israel T; Valieris, Renan et al. (2018) Clonal CD4+ T cells in the HIV-1 latent reservoir display a distinct gene profile upon reactivation. Nat Med 24:604-609 |
| Horwitz, Joshua A; Bar-On, Yotam; Lu, Ching-Lan et al. (2017) Non-neutralizing Antibodies Alter the Course of HIV-1 Infection In Vivo. Cell 170:637-648.e10 |
| Nishimura, Yoshiaki; Gautam, Rajeev; Chun, Tae-Wook et al. (2017) Early antibody therapy can induce long-lasting immunity to SHIV. Nature 543:559-563 |
| Bournazos, Stylianos; Ravetch, Jeffrey V (2017) Anti-retroviral antibody Fc?R-mediated effector functions. Immunol Rev 275:285-295 |
| Gristick, Harry B; Wang, Haoqing; Bjorkman, Pamela J (2017) X-ray and EM structures of a natively glycosylated HIV-1 envelope trimer. Acta Crystallogr D Struct Biol 73:822-828 |
| Bournazos, Stylianos; Wang, Taia T; Dahan, Rony et al. (2017) Signaling by Antibodies: Recent Progress. Annu Rev Immunol 35:285-311 |
| Bournazos, Stylianos; Ravetch, Jeffrey V (2017) Fc? Receptor Function and the Design of Vaccination Strategies. Immunity 47:224-233 |
Showing the most recent 10 out of 52 publications
