L-Arginine and L-Glutamic Acid Increase the Content of Protein C in the Early Stages of Isolation From Donor Plasma

Abstract

Current large-scale production of blood-derived pharmacological preparations is aimed at expanding the list of products and deeper extraction of target proteins especially at the pre-purification stage. In particular, this problem becomes critical for the isolation of proteins like protein C (PC), which is present in plasma in trace amounts. Aim. We aimed to improve the buffer composition to minimize the interaction of PC with other proteins and lipids that are inevitably present in the stock material. Methods. The content of protein C in plasma and its derivatives was assessed by the amidolytic activity to the chromogenic substrate S2366. A decrease in homologous impurities and plasma enrichment with protein C was provided by selective bulk adsorption on DEAE-cellulose. Results. Here we describe that an equimolar mixture of two amino acids (L-arginine and L-glutamic acid) essentially increased the content of protein C at the stage of cryo-depleted plasma pre-purification, including initial dilution and subsequent enrichment of plasma with protein C due to selective bulk adsorption on DEAE- cellulose. Additionally, it was revealed that solutions of these amino acids, when combined, inhibit the induced amidolytic activity of protein C and increase its solubility (in contrast to other plasma proteases). Conclusion. Pre-adding of a mixture of amino acids L-arginine and L-glutamic acid to cryo-depleted plasma significantly optimizes the pre-purification stage of protein C, providing a 5-fold increase in its yield after elution from DEAE-cellulose.

Аrginine – biological role, biosynthesis and whey consumption

“…В останні роки зростає виробництво L-аргініну у зв'язку із розширенням сфер його застосування, а саме, в медицині, косметології, фармацевтичній промисловості, розведенні та годівлі тварин, виробництві нутріцевтиків (Soeters et al, 2004;Oshimura et al, 2007;Xu et al, 2007;Suzuki, 2013;Patalakh et al, 2021;Ginésy, 2021).…”

Isolation of DNA from spots of old microscopic glass slides with mini column isolation kit for molecular analysis

“…Isolation of nucleic acid is an essential step for conducting the different nucleic acid analysis methods like PCR and hybridization for various viral, bacterial, biomarkers and genetic targets [1][2][3][4]. Not only this, but the nucleic acid can also be used as therapeutic as well as vaccine options in many clinical settings, therefore there is an urgent need to isolate the sufficient amount of nucleic acid in highly pure form [5][6][7][8] The nucleic acid can be isolated from different sources e.g., blood, tissue, buccal swabs, nasal samples, plasma, serum, urine and cell cultures.…”