Western blot assay for quantitative and qualitative antigen detection in vaccine development.
Immunological methods for quantitative measurement, antigenic characterization, and monitoring the stability of active immunogenic component(s) are a critical need in the vaccine development process. This unit describes an enhanced chemiluminescence-based western blot for quantitative detection of Plasmodium falciparum circumsporozoite protein (PfCSP), a major malaria candidate vaccine antigen. The most salient features of this assay are its high sensitivity and reproducibility; it can reliably detect ∼5 to 10 pg PfCSP expressed on native parasites or recombinantly expressed in Escherichia coli. Although described for a specific vaccine antigen, this assay should be applicable for any antigen-antibody combination for which relevant detection reagents are available. Detailed stepwise experimental procedures and methods for data acquisition and analysis are described.
Laboratory of Emerging Pathogens, Division of Emerging and Transfusion Transmitted Diseases, Office of Blood Research and Review, Center for Biologics Evaluation and Research, Food and Drug Administration, Rockville, Maryland.
Sanjai
S
Kumar
Hong
H
Zheng
Babita
B
Mahajan
Yukiko
Y
Kozakai
Merribeth
M
Morin
Emily
E
Locke
D000953:Antigens, Protozoan / Q000032:analysis* / Q000276:immunology; D015153:Blotting, Western / Q000379:methods*; D017780:Malaria Vaccines / Q000032:analysis* / Q000276:immunology; D010963:Plasmodium falciparum / Q000276:immunology*; D015800:Protozoan Proteins / Q000032:analysis* / Q000276:immunology; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D015203:Reproducibility of Results
D000953:Antigens, Protozoan; D017780:Malaria Vaccines; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C041595:circumsporozoite protein, Protozoan
antigen; malaria; stability; vaccine; western blot