title: A novel site-II directed glycoprotein estimation ELISA to aid rabies vaccine manufacture for veterinary and human use.
abstract: Although the World Health Organization recommends the use of in vitro techniques to qualify rabies vaccine lot release, very limited proposals have been made to arrive at a harmonized approach for wide scale usage. The present study proposed and evaluated the use of a novel avidin-biotin ELISA as an alternative to these in vivo tests in rabies vaccine manufacture. This assay utilized a neutralizing pan reactive monoclonal antibody (mAb) reactive with the conserved site-II of the natively folded rabies glycoprotein. Linear regression analysis of the in vitro glycoprotein estimates with the in vivo potency values, showed a good correlation (r(2)=0.8) with veterinary vaccines, but a poor correlation (r(2)=0.2) with human vaccines. However, we could qualitatively arrive at cut-off glycoprotein estimates from the ELISA, above which all the vaccines were declared to be protective by mouse challenge studies (>2.5IU/dose).
authors.affiliation: Indian Immunologicals Limited, Gachibowli, Hyderabad 500032, Andhra Pradesh, India.
authors.forename: Gontu
authors.initials: G
authors.lastname: Abhinay
authors.affiliation: Lankenau Institute for Medical Research, Wynnewood, PA 19096, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Dessain
authors.affiliation: Indian Immunologicals Limited, Gachibowli, Hyderabad 500032, Andhra Pradesh, India.
authors.forename: Adabala
authors.initials: A
authors.lastname: Srikanth
authors.affiliation: Indian Immunologicals Limited, Gachibowli, Hyderabad 500032, Andhra Pradesh, India.
authors.forename: R L
authors.initials: RL
authors.lastname: Senthilkumar
authors.affiliation: Indian Immunologicals Limited, Gachibowli, Hyderabad 500032, Andhra Pradesh, India.
authors.forename: Pitta
authors.initials: P
authors.lastname: Vidyasagar
authors.affiliation: Indian Immunologicals Limited, Gachibowli, Hyderabad 500032, Andhra Pradesh, India.
authors.forename: Alagangula
authors.initials: A
authors.lastname: Praveen
authors.affiliation: Indian Immunologicals Limited, Gachibowli, Hyderabad 500032, Andhra Pradesh, India.
authors.forename: R V
authors.initials: RV
authors.lastname: Chandrasekhar Reddy
authors.affiliation: Indian Immunologicals Limited, Gachibowli, Hyderabad 500032, Andhra Pradesh, India.
authors.forename: Erri
authors.initials: E
authors.lastname: Swapna Reddy
authors.affiliation: Indian Immunologicals Limited, Gachibowli, Hyderabad 500032, Andhra Pradesh, India. Electronic address: rlingala2005@gmail.com.
authors.forename: Lingala
authors.initials: L
authors.lastname: Rajendra
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D051379:Mice; D011818:Rabies / Q000517:prevention & control; D011819:Rabies Vaccines / Q000032:analysis*; D014764:Viral Proteins / Q000032:analysis*
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D006023:Glycoproteins; D011819:Rabies Vaccines; D014764:Viral Proteins
keywords: Antigenic site-II; Avidin–biotin complex ELISA; Glycoprotein; Mouse potency; Rabies vaccines