Use of o-phthalaldehyde assay to determine protein contents of Alhydrogel-based vaccines.

Abstract:

:Aluminum based adjuvants (alum), including aluminum hydroxide (Alhydrogel) and aluminum phosphate are the most commonly used adjuvant in the US. In order to ensure quality of vaccines, regulatory authorities require evaluation of antigen content in final vaccine products. Currently, there are no generic methods available for the determination of protein content in alum-based vaccines. Aluminum hydroxide gels exist as particles in solution, which interfere with direct quantitation of protein content in formulations using assays such as Lowry, BCA or Bradford protein assay. The present study adapts a simple fluorescent assay to directly (without the need for antigen extraction) determine antigen content on Alhydrogel with accuracy and sensitivity using the o-phthalaldehyde (OPA) reagent. Malaria vaccine candidates AMA1-C1/Alhydrogel, AMA1-C2/Alhydrogel, MSP1(42)-3D7/Alhydrogel, MSP1(42)-C1/Alhydrogel or BSAM-2/Alhydrogel were used as model formulations. The results of the present study show that the OPA assay is highly accurate (87-100%), reproducible, and simple with a linear detection range of 25-400 microg/mL for Alhydrogel vaccines (except for MSP1(42)-C1, which has a linear detection range of 31.25-500 microg/mL). This assay has proven to be highly useful in our laboratory and been used in routine vaccine quality control processes.

journal_name

Vaccine

journal_title

Vaccine

authors

Zhu D,Saul A,Huang S,Martin LB,Miller LH,Rausch KM

doi

10.1016/j.vaccine.2009.07.067

subject

Has Abstract

pub_date

2009-10-09 00:00:00

pages

6054-9

issue

43

eissn

0264-410X

issn

1873-2518

pii

S0264-410X(09)01090-1

journal_volume

27

pub_type

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