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Electron-Microscopy-Based Epitope Mapping Defines Specificities of Polyclonal Antibodies Elicited during HIV-1 BG505 Envelope Trimer Immunization.

Immunity. 2018;

Bianchi M, Turner HL, Nogal B, Cottrell CA, Oyen D, Pauthner M, Bastidas R, Nedellec R, McCoy LE, Wilson IA, Burton DR, Ward AB, Hangartner L.

Products/Services Used	Details	Operation
Peptide Synthesis	Competition with BG505 V3-peptide (TRPNNNTRKSIRIGPGQAFYATGDIIGDIRQAH, GenScript) was performed by pre-incubation of antibodies or plasmas with 150 mg/mL V3-peptide at room temperature for 1 hr before incubation on BG505 SOSIP.... Biotinylated BG505 V3-peptide (TRPNNNTRKSIRIGPG QAFYATGDIIGDIRQAH, GenScript) was captured on the neutravidin plate for 2 hr at RT, before adding serial dilutions of Fab or plasma for additional 2 hr at RT.	Get A Quote

摘要

Characterizing polyclonal antibody responses via currently available methods is inherently complex and difficult. Mapping epitopes in an immune response is typically incomplete, which creates a barrier to fully understanding the humoral response to antigens and hinders rational vaccine design efforts. Here, we describe a method of characterizing polyclonal responses by using electron microscopy, and we applied this method to the immunization of rabbits with an HIV-1 envelope glycoprotein vaccine candidate, BG505 SOSIP.664. We detected known epitopes within the polyclonal sera and revealed how antibody responses evolved during the prime-boosting strategy to ultimately result in a neutralizing antibody response. ... More