Validation of the modified Lowry to quantify the protein content of an allergy vaccine

Abstract

Introduction: Prolinem BT is a second-generation vaccine candidate that uses as active ingredient allergens purified from Blomia tropicalis mite extract, which constitute the major allergens of that species.

Objective: Validation of the microplate modified Lowry as a technique to quantify the total protein content of the active ingredient of the PROLINEM-BT vaccine.

Methods: For the quantification of these purified proteins, the microplate modified Lowry technique was developed and validated. The parameters studied in the validation were specificity, linearity and parallelism, precision and accuracy.

Results: The method was specific in the quantification of total proteins. The curve ranges from 20 to 100 µg/mL and had a linear behavior (r = 0.99). The technique is precise based on the coefficients of variation obtained (all less than 5 %) and accurate since all values presented 100 % recovery.

Conclusions: It was demonstrated that the Lowry method modified to microplates is valid for the quantification of total proteins of the active ingredient of Prolinem BT.