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HomeWebinarsA Sensitive Method to Quantify HIV-1 Antibody in Mucosal Samples

A Sensitive Method to Quantify HIV-1 Antibody in Mucosal Samples

Webinar

The AIDS epidemic is a global challenge with 38 million people living with HIV and 1.7 million new infections along with 690K deaths in 2019. To date, HIV-1 vaccine clinical trials have not been successful in eliciting robust broad neutralizing antibody responses that can prevent infections. Alternative approaches, such as passive immunization with broadly neutralizing monoclonal antibodies, are being considered to help prevent new HIV infections. VRC01, an HIV-gp120 CD4bs directed potent and broadly neutralizing antibody (bNAb), which neutralizes about 91% of HIV-1 strains of a panel consisting of 190 Env-pseudotyped viral strains with a half max inhibitory concentration (IC50) of <50 ug/mL, is being tested for safety and efficacy in preventing HIV infection. Since the majority of HIV-1 transmissions occur through mucosal routes, it is essential to quantify the levels of the passively administered bNAb in mucosal samples. This helps to determine the levels of the bNAbs that have localized to mucosal sites preventing infection by neutralizing the virus. Traditional ELISAs and other immunoassays with a lower limit of quantification (LLOQ) in the range of 1 ug/mL are not sensitive enough to accurately quantify the low levels of VRC01 in mucosal samples. Hence, we sought to develop a custom ultra-sensitive Single Molecule Counting (SMC™) magnetic microparticle-based immunoassay with a greater than 4-log linear dynamic range (LDR) and LLOQ less than 120 pg/mL. We successfully quantified VRC01 levels in rectal, cervical and oral mucosal samples in two passive immunization studies conducted with VRC01- VRC 601 and VRC 602. The assay was able to successfully quantify VRC01 levels in mucosal samples from all dosage groups from these trials. This accurate and sensitive immunoassay can be utilized in vaccine research for quantifying low antibody levels in mucosal and other sample types in the context of HIV-1 pre-clinical and clinical vaccine trials.

Speakers

Sandeep Narpala

Vaccine Research Center (VRC), NIAID, NIH

Scientific Project Manager, Vaccine Immunology Program (VIP)

Sandeep Narpala is a Scientific Project Manager at the Vaccine Immunology Program (VIP), part of the Vaccine Research Center (VRC) within the National Institute of Allergy and Infectious Diseases (NIAID) at the National Institutes of Health (NIH). Sandeep manages the procurement of reagents, development, optimization, and qualification of novel clinical immunoassays and schedules the testing of pre-clinical and clinical trial samples conducted at VRC. Mr. Narpala has been affiliated with the VRC/NIH since 2011 and has extensive knowledge in assay development for vaccine research on various platforms such as Meso Scale Discovery (MSD), Single Molecule Counting (SMC™) technology, and other ELISA-based platforms. Sandeep has successfully developed and qualified immunogenicity assays to assess secondary objectives in HIV and influenza vaccine clinical trials and has expertise in the development of pharmacokinetic and anti-drug antibodies assays for passive monoclonal antibody trials conducted at VRC. Before working at NIH, Mr. Narpala honed his skills working as a biologist at International Aids Vaccine Initiative (IAVI) in New York City. With his bioinformatics background, Sandeep was involved in the in-silico design of HIV immunogens using Molecular Dynamics simulations at IAVI. Sandeep has a Masters in Bioinformatics and graduated from Northeastern University, Boston in 2009. Sandeep plans to grow as a Project Manager at VIP and is currently involved in managing and overseeing the development of immunoassays to screen new COVID-19 spike specific antibodies and multiplexed assays to detect binding COVID-19 antibodies in sero surveillance studies.

Webinar Information

Research Areas

  • Immunology
  • Duration:41 mins

  • Language:English

  • Presented: Wed, March 31, 2021