Quantitative Analyses of all Influenza Type A Viral Hemagglutinins and Neuraminidases using Universal Antibodies in Simple Slot Blot Assays

Caroline Gravel; Changgui Li; Junzhi Wang; Anwar M Hashem; Bozena Jaentschke; Gary Van Domselaar; Runtao He; Xuguang Li

doi:10.3791/2784

Quantitative Analyses of all Influenza Type A Viral Hemagglutinins and Neuraminidases using Unive...

JoVE Journal
Immunology and Infection

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Immunology and Infection

Quantitative Analyses of all Influenza Type A Viral Hemagglutinins and Neuraminidases using Universal Antibodies in Simple Slot Blot Assays

Full Text

18,798 Views

13:12 min

April 4, 2011

DOI: 10.3791/2784-v

Caroline Gravel*¹, Changgui Li*², Junzhi Wang², Anwar M Hashem^1,3,4, Bozena Jaentschke¹, Gary Van Domselaar⁵, Runtao He⁵, Xuguang Li^1,3

¹Centre for Vaccine Evaluation, Biologics and Genetic Therapies Directorate, HPFB,Health canada, ²National Institute for the Control of Pharmaceutical and Biological Products,The State Food and Drug Administration, Beijing, ³Department of Biochemistry, Microbiology and Immunology,University of Ottawa, ⁴Microbiology Department, Faculty of Medicine,King Abdulaziz University, ⁵National Microbiology Laboratory,Public Health Agency of Canada

A simple slot blot method was developed for the quantification of influenza viral hemagglutinin and neuraminidase using universal antibodies targeting their most conserved sequences identified through bioinformatics analyses. This innovative approach may provide a useful alternative to quantitative determination of all viral hemagglutinin and neuraminidase.

The overall goal of this procedure is to quantitatively determine the influenza viral hemagglutinin and neuraminidase content in vaccines using antibodies targeting conserved epitopes. This is accomplished by first identifying conserved epitopes. The second step of the procedure is to dilute the reference and vaccine samples to improve the detection sensitivity.

The third step is to assemble the slot blot apparatus and load the samples into the slots. The fourth step of the procedure is to detect the HA or NA on the membrane using their respective universal antibodies. The final step of the procedure is to quantify the amounts of HA or NA by densitometry.

View the full transcript and gain access to thousands of scientific videos