Quantifying Vibriocidal Antibodies against Cholera

Murine and human vibriocidal antibodies were quantified by finding the highest serum dilution required to lyse isogenic ZChol strains or PIC158 (Ogawa) or PIC018 (Inaba) V. cholerae as described previously (28 (link), 62 (link)). Heat-inactivated 2-fold serial dilutions of serum were incubated with guinea pig complement (Sigma) and the target strain and then allowed to grow in brain heart infusion (BHI) medium in a 96-well plate. The serum dilution that caused more than 50% reduction in target strain OD₅₉₅ compared to saline negative-control wells was recorded as the antibody titer. A mouse monoclonal antibody, 432A.1G8.G1.H12 (28 (link)), targeting V. cholerae O1 OSP was a positive control for the assay. The limit of detection represents the lowest serum dilution at which no inhibition of growth could be detected. The r² value was determined using a linear regression model in Prism 9 (GraphPad), and the data were represented on a log₁₀ scale for ease of interpretation.

Free full text: Click here

Fakoya B., Hullahalli K., Rubin D.H., Leitner D.R., Chilengi R., Sack D.A, & Waldor M.K. (2022). Nontoxigenic Vibrio cholerae Challenge Strains for Evaluating Vaccine Efficacy and Inferring Mechanisms of Protection. mBio, 13(2), e00539-22.

Publication 2022

guinea pig complement Prism 9

Antibodies Antibody Assay Brain Dilution Guinea pig Heart Human Inhibition Monoclonal antibody Mouse Murine Prism Saline Serum Strain V cholerae

Corresponding Organization : Howard Hughes Medical Institute

Other organizations : Centre for Infectious Disease Research in Zambia, Johns Hopkins University

Top 5 similar protocols

Variable analysis

independent variables

Serum dilution

dependent variables

Vibriocidal antibody titer

control variables

Incubation with guinea pig complement
Growth in brain heart infusion (BHI) medium
Saline negative-control wells

controls

Positive control: Mouse monoclonal antibody 432A.1G8.G1.H12 targeting V. cholerae O1 OSP

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!