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Statistical optimization of immunoaffinity purification of hepatitis B surface antigen using response surface methodology

Paper ID Volume ID Publish Year Pages File Format Full-Text
4734 241 2008 8 PDF Available
Title
Statistical optimization of immunoaffinity purification of hepatitis B surface antigen using response surface methodology
Abstract

Response surface methodology (RSM) was successfully applied to find an elution condition enhancing the performance of immunoaffinity (IAF) purification of hepatitis B surface antigen (HBsAg), conducted in batch chromatography. The immunosorbent efficiency measured as HBsAg eluted per milliliter of immunosorbent was selected as the response. Two numerical independent variables were studied: concentration of potassium thiocyanate (KSCN) and ethanol, and a categorical variable formed by two levels of sodium deoxycholate under defined ionic strength (DCS/NaCl). A three-level factorial design of RSM was developed and gets a reduced cubic model which accurately fits the data (R2 ≈ 0.97). RSM predicted an optimal region where the response could allow maximum values. At the optimal condition 1.7 M KSCN, 30% (v/v) ethanol, and 0% (w/v) DCS and 1 M NaCl a response of 582 ± 11 μg/ml (559 ± 52 by the model), wherein the measured response does not differs statistically with the maximum experimental value. At optimized condition, a recovery of HBsAg of ∼94% of the adsorbed antigen was obtained, representing a ∼69% more HBsAg than the previously reported. Due to less accumulation of HBsAg this optimum could increase the immunosorbent life in a multiple cycle's operation.

Keywords
ANOVA, analysis of variance; API, active pharmaceutical ingredient; CI, confidence interval; CDR, complementary determining region; CV, coefficient of variations; DCS, sodium deoxycholate; ELISA, enzyme-linked immunosorbent assay; EtOH, ethanol; HBsAg, he
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Statistical optimization of immunoaffinity purification of hepatitis B surface antigen using response surface methodology
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Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 38, Issue 1, 15 January 2008, Pages 1–8
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
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Any Questions? feel free to contact us