title: Trypsin diminishes the rat potency of polio serotype 3.
abstract: This study addresses observations made in view of testing in practice the guideline in the European Pharmacopoeia (EP) on omitting the rat potency test for release of polio containing vaccines. In general, use of the guideline is valid and the D-antigen ELISA can indeed be used as an in vitro alternative for the in vivo test. However, the set-up of the ELISA is crucial and should include detection of antigenic site 1 in polio serotype 3 as destruction of that site by trypsin results in a reduced rat potency. Antigenic site 1 in polio serotype 2 may also be modified by trypsin, but the cleavage of viral protein 1 (VP1) did not affect the rat potency. Therefore, any antigenic site, except site 1, can be used for detection of polio serotype 2. It is advised to include testing of the effect of trypsin treatment in the EP-guideline. This allows polio vaccine manufacturers to check whether their in-house ELISA needs improvement.
authors: Institute for Translational Vaccinology, P.O. Box 450, 3720 AL Bilthoven, The Netherlands. Electronic address: Rimko.ten.Have@intravacc.nl.
authors: R
authors: R
authors: ten Have
authors: Institute for Translational Vaccinology, P.O. Box 450, 3720 AL Bilthoven, The Netherlands.
authors: J
authors: J
authors: Westdijk
authors: Bilthoven Biologicals, P.O. Box 457, 3720 AL Bilthoven, The Netherlands.
authors: L M A R
authors: LM
authors: Levels
authors: Bilthoven Biologicals, P.O. Box 457, 3720 AL Bilthoven, The Netherlands.
authors: P
authors: P
authors: Koedam
authors: Institute for Translational Vaccinology, P.O. Box 450, 3720 AL Bilthoven, The Netherlands.
authors: A
authors: A
authors: de Haan
authors: Institute for Translational Vaccinology, P.O. Box 450, 3720 AL Bilthoven, The Netherlands.
authors: M R J
authors: MR
authors: Hamzink
authors: Institute for Translational Vaccinology, P.O. Box 450, 3720 AL Bilthoven, The Netherlands.
authors: B
authors: B
authors: Metz
authors: Institute for Translational Vaccinology, P.O. Box 450, 3720 AL Bilthoven, The Netherlands; Leiden Academic Center for Drug Research, Drug Delivery Technology, P.O. Box 9502, 2300 RA Leiden, The Netherlands.
authors: G F A
authors: GF
authors: Kersten
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000096:biosynthesis; D000914:Antibodies, Viral / Q000096:biosynthesis; D001681:Biological Assay; D015374:Biosensing Techniques; D036022:Capsid Proteins / Q000187:drug effects; D002522:Chlorocebus aethiops; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D005060:Europe; D006358:Hot Temperature; D017955:Poliovirus / Q000187:drug effects* / Q000276:immunology / Q000302:isolation & purification / Q000472:pathogenicity; D011054:Poliovirus Vaccine, Inactivated / Q000276:immunology*; D017410:Practice Guidelines as Topic; D051381:Rats; D014357:Trypsin / Q000494:pharmacology*; D014709:Vero Cells; D014774:Virulence / Q000187:drug effects
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D036022:Capsid Proteins; D011054:Poliovirus Vaccine, Inactivated; C442540:VP1 protein, Poliovirus; D014357:Trypsin
keywords: D-antigen; ELISA; Inactivated polio vaccine; Rat potency test; Sub-potent; Test animals