Another paper on anti-glycophorin-A MAb "purification" (compare to the one described in the post "Acid Test for antibody function?" by Ambur P. Dhivya's 2010 paper in Chromatographia)
CIM-EDA has been used in both papers. In this latest one (2012), ~ 8 fold purification was seen with elution with 25 mM MOPS, pH 6.5 and 0.2 M NaCl.
In the older one (2010), 20 mM Tris buffer, pH 8, 0.6 M NaCl gave ~74 fold purification.
Images are still difficult to make out; but why change the conditions and report worse "purification" with different conditions?
Side-by-side comparisons of chromatographic purification gels and profiles from the older (blue arrows and underlined in blue) Chromatographia 2010 paper and the recent (red arrows and underlined in red) Hybridoma 2012 paper.
Kumar BP, Rajak P, Vijayalakshmi MA, & Jayaprakash NS (2012). Production of Human Anti-Glycophorin-A Monoclonal Antibodies and Their Purification by Pseudoaffinity Chromatography Using a Convective Interaction Media Monolithic Column Hybridoma DOI: 10.1089/hyb.2011.0104
Dhivya AP, Kumar BP, Prasanna RR, Jayaprakash NS, & Vijayalakshmi MA (2010). Purification of Monoclonal Antibodies from Cell-Culture Supernatant by Use of Anion-Exchange Convective Interaction Media (CIM) Monolithic Columns Chromatographia, 72 (11-12) DOI: 10.1365/s10337-010-1787-3
CIM-EDA has been used in both papers. In this latest one (2012), ~ 8 fold purification was seen with elution with 25 mM MOPS, pH 6.5 and 0.2 M NaCl.
In the older one (2010), 20 mM Tris buffer, pH 8, 0.6 M NaCl gave ~74 fold purification.
Images are still difficult to make out; but why change the conditions and report worse "purification" with different conditions?
abstract from Ambur Dhivya et al. 2010 Chromatographia
Dead cells producing antibody?
Side-by-side comparisons of chromatographic purification gels and profiles from the older (blue arrows and underlined in blue) Chromatographia 2010 paper and the recent (red arrows and underlined in red) Hybridoma 2012 paper. Kumar BP, Rajak P, Vijayalakshmi MA, & Jayaprakash NS (2012). Production of Human Anti-Glycophorin-A Monoclonal Antibodies and Their Purification by Pseudoaffinity Chromatography Using a Convective Interaction Media Monolithic Column Hybridoma DOI: 10.1089/hyb.2011.0104
Dhivya AP, Kumar BP, Prasanna RR, Jayaprakash NS, & Vijayalakshmi MA (2010). Purification of Monoclonal Antibodies from Cell-Culture Supernatant by Use of Anion-Exchange Convective Interaction Media (CIM) Monolithic Columns Chromatographia, 72 (11-12) DOI: 10.1365/s10337-010-1787-3
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