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Title: PROTEIN AND PEPTIDE DETERMINATION BASED ON THE MODIFIED BIURET PROCEDURE: IMPLICATIONS FOR VARIOUS BIOTECHNOLOGIES
PROTEIN AND PEPTIDE DETERMINATION BASED ON THE MODIFIED BIURET PROCEDURE: IMPLICATIONS FOR VARIOUS BIOTECHNOLOGIES

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Elena Mihalcea; Gabi Drochioiu; Stefania-Claudia Jitaru; Violeta Mangalagiu; Robert –Vasile Gradinaru
10.5593/sgem2022/6.1/s25.14
10.5593/sgem2022/6.1
1314-2704
978-619-7603-48-4
English
22
6.1
•    Prof. DSc. Oleksandr Trofymchuk, UKRAINE 
•    Prof. Dr. hab. oec. Baiba Rivza, LATVIA
Advances in Biotechnology
Spectrophotometric methods for total protein analysis are generally simple, rapid and sensitive. Such sensitive protein assays may have applications in forensic science, in the detection of protein contaminants in drugs and in a number of other applications of research interest. Biuret reaction with proteins and peptides is widely used in clinical and biological laboratories. In this work, instead of copper sulphate, sodium hydroxide and Seignette salt, we used insoluble copper phosphate, potassium or sodium hydroxide and ethyl alcohol for the preparation of the biuret reagent. Absorbance of the biuret complex was recorded both at 219-230 nm (after dilution) and around 550 nm against a reagent blank. Amino acid interference was investigated around 550 nm at the same concentration as proteins. The sensitivity of the method at 226 nm was greater than those of other spectrophotometric assays (old biuret method, Lowry, and BCA) with a LOD of about 0.5 µg mL?1 BSA. The new variants of the biuret method for total protein analysis eliminate the need for precise reagent addition and vortexing inherent in the widely used Lowry method, providing flexibility of application. The method developed, which uses an alkaline-alcoholic reagent and insoluble copper phosphate, is simple, rapid, reproducible and sensitive; it is not influenced by detergents, solvents and buffers containing ammonium and is flexible enough to change the analytical protocol when necessary. A discussion was made on the applications of protein and peptide determination with the new biuret assay.
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Financial support from the Romanian Government through UEFISCDI Bucharest (PNIII-P2-2.1-PED2019-2484, Contract PED494, BioPASCAL G.D. also acknowledges Contract no.11PFE/30.12.2021 from Romanian Ministry of Research, Innovation and Digitization.
conference
https://doi.org/10.5593/sgem2022/6.1/s25.14
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Proceedings of 22nd International Multidisciplinary Scientific GeoConference SGEM 2022
22nd International Multidisciplinary Scientific GeoConference SGEM 2022, 04 - 10 July, 2022
Proceedings Paper
STEF92 Technology
International Multidisciplinary Scientific GeoConference SGEM
SWS Scholarly Society; Acad Sci Czech Republ; Latvian Acad Sci; Polish Acad Sci; Serbian Acad Sci and Arts; Natl Acad Sci Ukraine; Natl Acad Sci Armenia; Sci Council Japan; European Acad Sci, Arts and Letters; Acad Fine Arts Zagreb Croatia; Croatian Acad Sci and Arts; Acad Sci Moldova; Montenegrin Acad Sci and Arts; Georgian Acad Sci; Acad Fine Arts and Design Bratislava; Turkish Acad Sci.
113-120
04 - 10 July, 2022
website
8642
Protein determination, Peptide determination, Biuret reaction, Amino acid interference, Spectrophotometry