High Enrichment of MMP-9 and Carboxypeptidase A by Tweezing Adsorptive Bubble Separation (TABS)

Abstract

Tweezing adsorptive bubble separation (TABS) was used as a method for the enrichment of matrix metalloproteinases (92-kDa type IV, gelatinase B (MMP-9)) and carboxypeptidase A (CPA) from dilute aqueous solutions. The method is based on the chelation of metalloenzymes applying 2-(carbamoylmethyl-(carboxymethyl)amino)acetic acid (ADA) coupled with an octyl part to form a surface active unit. MMP-9 could be enriched with an enrichment ratio of 12.0 and a recovery of 87.3%, and CPA could be enriched 18.8-fold and with 95.3% recovery. Both enzymes were enriched without significant losses of enzymatic activity. To verify that the enzymes were tweezed by ADA-C8 without abstraction of the zinc ions from the active center, TABS trials were additionally conducted with zinc ions in complex with ADA-C8, which revealed only negligible enrichment ratios of the enzymes (2.2 for MMP-9 and 0.2 for CPA). The results obtained impressively demonstrate that zinc-containing proteases can be enriched selectively and efficiently by TABS.

Enrichment behavior of immunoglobulin by foam fractionation using response surface methodology

“…The current interest seems to indicate a desire for a low-cost, highly effective purification method for biological products, particularly in the first concentration step that is used to remove about 90% of the water from dilute solutions. Several papers further support these studies by providing a theoretical understanding and empirical observations of protein foam fractionation [HALLER, et al 2010, LINKE, et al 2007, LOCKWOOD, et al 2000, URAIZEE and NARSIMHAN 1990a, URAIZEE and NARSIMHAN 1990b.…”