65A-12: Reductive alkylation of blood proteins

RE Nyman, G Nelson, R Owusu-Apenten

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Globin and plasma were chemically modified by reductive alkylation using 4-hydroxybenzaldehyde and the reducing agent sodium cyanoborohydride. The resulting derivatives were analysed to determine their solubility and protein concentration. The degree of modification was estimated by amino acid analysis and sodium dodecyl-sulphate polyacrylamide gel electrophoresis. The protein solutions were brought to neutrality with sodium hydroxide to allow optimum enzymatic cross-linking. RESULTS: The derivatives were found to be very insoluble at pH 6; optimum solubility (90-95%) was found at pH 2 and 60?C (pre-heating). The optimum pH values for tyrosinase-catalysed cross-linking of globin and plasma derivatives were 7 and 8, respectively. Kjeldhal nitrogen analysis resulted in estimates of the protein content of the derivatives at approximately 75%. Analysis of the derivatives proved that modification had occurred. Amino acid analysis determined that globin and plasma were 73% and 27% modified, respectively. Electrophoresis confirmed a qualitative increase in molecular weight. SIGNIFICANCE: Enzymatic cross-linking of proteins is an alternative and potentially less expensive method of gelation, carried out at low temperature compared to the traditional heat-induced method
LanguageEnglish
Title of host publicationUnknown Host Publication
Place of PublicationChicago, IL
Number of pages1
Publication statusPublished - 1999
Event1999 IFT Annual Meeting, July 24 - 28, Chicago, IL -
Duration: 1 Jan 1999 → …

Conference

Conference1999 IFT Annual Meeting, July 24 - 28, Chicago, IL
Period1/01/99 → …

Fingerprint

alkylation
blood proteins
chemical derivatives
crosslinking
solubility
heat
amino acids
reducing agents
proteins
sodium hydroxide
gelation
electrophoresis
polyacrylamide gel electrophoresis
protein content
sodium
molecular weight
nitrogen
methodology
temperature

Cite this

Nyman, RE., Nelson, G., & Owusu-Apenten, R. (1999). 65A-12: Reductive alkylation of blood proteins. In Unknown Host Publication Chicago, IL.
Nyman, RE ; Nelson, G ; Owusu-Apenten, R. / 65A-12: Reductive alkylation of blood proteins. Unknown Host Publication. Chicago, IL, 1999.
@inproceedings{e113e657ae9c4171a5a52e6c732b509e,
title = "65A-12: Reductive alkylation of blood proteins",
abstract = "Globin and plasma were chemically modified by reductive alkylation using 4-hydroxybenzaldehyde and the reducing agent sodium cyanoborohydride. The resulting derivatives were analysed to determine their solubility and protein concentration. The degree of modification was estimated by amino acid analysis and sodium dodecyl-sulphate polyacrylamide gel electrophoresis. The protein solutions were brought to neutrality with sodium hydroxide to allow optimum enzymatic cross-linking. RESULTS: The derivatives were found to be very insoluble at pH 6; optimum solubility (90-95{\%}) was found at pH 2 and 60?C (pre-heating). The optimum pH values for tyrosinase-catalysed cross-linking of globin and plasma derivatives were 7 and 8, respectively. Kjeldhal nitrogen analysis resulted in estimates of the protein content of the derivatives at approximately 75{\%}. Analysis of the derivatives proved that modification had occurred. Amino acid analysis determined that globin and plasma were 73{\%} and 27{\%} modified, respectively. Electrophoresis confirmed a qualitative increase in molecular weight. SIGNIFICANCE: Enzymatic cross-linking of proteins is an alternative and potentially less expensive method of gelation, carried out at low temperature compared to the traditional heat-induced method",
author = "RE Nyman and G Nelson and R Owusu-Apenten",
year = "1999",
language = "English",
booktitle = "Unknown Host Publication",

}

Nyman, RE, Nelson, G & Owusu-Apenten, R 1999, 65A-12: Reductive alkylation of blood proteins. in Unknown Host Publication. Chicago, IL, 1999 IFT Annual Meeting, July 24 - 28, Chicago, IL, 1/01/99.

65A-12: Reductive alkylation of blood proteins. / Nyman, RE; Nelson, G; Owusu-Apenten, R.

Unknown Host Publication. Chicago, IL, 1999.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

TY - GEN

T1 - 65A-12: Reductive alkylation of blood proteins

AU - Nyman, RE

AU - Nelson, G

AU - Owusu-Apenten, R

PY - 1999

Y1 - 1999

N2 - Globin and plasma were chemically modified by reductive alkylation using 4-hydroxybenzaldehyde and the reducing agent sodium cyanoborohydride. The resulting derivatives were analysed to determine their solubility and protein concentration. The degree of modification was estimated by amino acid analysis and sodium dodecyl-sulphate polyacrylamide gel electrophoresis. The protein solutions were brought to neutrality with sodium hydroxide to allow optimum enzymatic cross-linking. RESULTS: The derivatives were found to be very insoluble at pH 6; optimum solubility (90-95%) was found at pH 2 and 60?C (pre-heating). The optimum pH values for tyrosinase-catalysed cross-linking of globin and plasma derivatives were 7 and 8, respectively. Kjeldhal nitrogen analysis resulted in estimates of the protein content of the derivatives at approximately 75%. Analysis of the derivatives proved that modification had occurred. Amino acid analysis determined that globin and plasma were 73% and 27% modified, respectively. Electrophoresis confirmed a qualitative increase in molecular weight. SIGNIFICANCE: Enzymatic cross-linking of proteins is an alternative and potentially less expensive method of gelation, carried out at low temperature compared to the traditional heat-induced method

AB - Globin and plasma were chemically modified by reductive alkylation using 4-hydroxybenzaldehyde and the reducing agent sodium cyanoborohydride. The resulting derivatives were analysed to determine their solubility and protein concentration. The degree of modification was estimated by amino acid analysis and sodium dodecyl-sulphate polyacrylamide gel electrophoresis. The protein solutions were brought to neutrality with sodium hydroxide to allow optimum enzymatic cross-linking. RESULTS: The derivatives were found to be very insoluble at pH 6; optimum solubility (90-95%) was found at pH 2 and 60?C (pre-heating). The optimum pH values for tyrosinase-catalysed cross-linking of globin and plasma derivatives were 7 and 8, respectively. Kjeldhal nitrogen analysis resulted in estimates of the protein content of the derivatives at approximately 75%. Analysis of the derivatives proved that modification had occurred. Amino acid analysis determined that globin and plasma were 73% and 27% modified, respectively. Electrophoresis confirmed a qualitative increase in molecular weight. SIGNIFICANCE: Enzymatic cross-linking of proteins is an alternative and potentially less expensive method of gelation, carried out at low temperature compared to the traditional heat-induced method

M3 - Conference contribution

BT - Unknown Host Publication

CY - Chicago, IL

ER -

Nyman RE, Nelson G, Owusu-Apenten R. 65A-12: Reductive alkylation of blood proteins. In Unknown Host Publication. Chicago, IL. 1999