Development of bead-based immunoassay to quantify neutralizing antibody for human papillomavirus 16 and 18.
Human papillomavirus (HPV) has drawn great attention globally because of its association with virtually all (99 %) cases of cervical cancer. HPV virus-like particles (VLPs) have been implicated as an effective HPV vaccine candidate. In this study, we optimized the relevant parameters for bacterial production of high-risk HPV16 and HPV18 VLP L1 proteins. The combination of glutathione S-transferase fusion and late log phase culture induction enhanced the solubility and yield of HPV L1 proteins. For detection and quantification of HPV-16 and -18 antibodies, a Luminex-based competitive immunoassay was developed for use in vaccine clinical trials. The characteristics of the assay that were optimized included monoclonal antibody specificity, conjugation of VLP to microspheres, VLP concentration, antibody concentration, dilution of samples, and incubation time. No cross-reactivity occurred. This immunoassay was proven to be sensitive and accurate, and is potentially valuable for vaccine candidate evaluation and clinical use.
Catholic Research Institute of Medical Science, Catholic University of Korea, Seoul, Republic of Korea.
Hai-Bo
HB
Liu
Pankaj Kumar
PK
Chaturvedi
Gantumur
G
Battogtokh
Hyo Joo
HJ
Bang
In-Wook
IW
Kim
Yong-Wan
YW
Kim
Kye-Shin
KS
Park
Woong Shick
WS
Ahn
D000911:Antibodies, Monoclonal / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D003429:Cross Reactions / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005982:Glutathione Transferase / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D052162:Human papillomavirus 16 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D052161:Human papillomavirus 18 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D030361:Papillomavirus Infections / Q000276:immunology / Q000517:prevention & control; D053918:Papillomavirus Vaccines / Q000276:immunology; D020673:Protein Renaturation; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D036022:Capsid Proteins; C114580:HPV L1 protein, Human papillomavirus; D009856:Oncogene Proteins, Viral; D053918:Papillomavirus Vaccines; D011994:Recombinant Proteins; D005982:Glutathione Transferase