JoVE Logo

로그인

JoVE 비디오를 활용하시려면 도서관을 통한 기관 구독이 필요합니다.

A Microneutralization Assay to Measure Neutralizing Antibody Titers in Human Serum

-- views • 1:34 min

내레이션 대본

For a microneutralization assay, take a multi-well plate containing heat-inactivated, serially diluted human serum with decreasing concentrations of influenza-specific neutralizing antibodies.

Introduce pathogenic influenza viruses. Serum antibodies interact with the viruses and neutralize them; however, higher serum dilutions exhibit limited virus neutralization.

Seed with Madine-Darby Canine Kidney cells, or MDCK cells, overexpressing human sialic acid residues on their surface.

In the higher dilution, non-neutralized influenza viruses fuse with the cells' sialic acid, releasing the viral genome into the cell cytoplasm.

The viral genes are transcribed and translated to viral proteins and nucleoproteins within the host cell.

Treat with a high concentration of acetone for fixation and permeabilization.

Introduce primary antibodies that interact with viral nucleoproteins.

Overlay with enzyme-conjugated secondary antibodies that interact with primary antibodies. Wash and treat with the enzyme's substrate, producing a color product.

Quantify the color intensities and plot them against the serum dilution; the highest serum dilution, achieving fifty percent virus neutralization represents the neutralization titer.

article

04:00

A Microneutralization Assay to Measure Neutralizing Antibody Titers in Human Serum

관련 동영상

47 Views

JoVE Logo

개인 정보 보호

이용 약관

정책

연구

교육

JoVE 소개

Copyright © 2025 MyJoVE Corporation. 판권 소유