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Comparing methods for immobilizing HIV-1 SOSIPs in ELISAs that evaluate antibody binding
Kim-Marie A Dam1, Patricia S Mutia2, Pamela J Bjorkman3
1Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA.
Scientific Reports
|July 1, 2022
Summary
Directly coating HIV-1 SOSIPs in ELISAs can alter Env structure, exposing hidden epitopes. This method may yield misleading results for broadly neutralizing antibody binding assays.
Area of Science:
- Immunology
- Virology
- Biochemistry
Background:
- Enzyme-linked immunosorbent assays (ELISAs) are crucial for assessing antibody binding to HIV-1 Env SOSIPs.
- Variability in antigen immobilization methods can impact ELISA results and antibody binding assessments.
Purpose of the Study:
- To investigate how different antigen immobilization methods affect HIV-1 Env SOSIP conformation.
- To determine the impact of immobilization techniques on the binding of broadly neutralizing antibodies (bNAbs).
Main Methods:
- Compared direct coating, antibody capture, and random biotinylation for immobilizing HIV-1 SOSIPs.
- Assessed binding of bNAbs targeting various epitopes using Enzyme-linked immunosorbent assays (ELISAs).
Main Results:
- Binding of most bNAbs was comparable across all immobilization methods.
- Direct coating led to detectable binding of the CD4-induced antibody 17b, unlike other methods, indicating altered Env conformation.
Conclusions:
- Direct coating of HIV-1 SOSIPs can disrupt Env structure, exposing cryptic epitopes.
- Antigen immobilization strategy significantly influences ELISA outcomes and the interpretation of antibody binding data.
