Improved immobilization of fusion proteins via cellulose-binding domains

Markus Linder (Corresponding Author), Tarja Nevanen, Lotta Söderholm, Outi Bengs, Tuula Teeri

Research output: Contribution to journalArticleScientificpeer-review

42 Citations (Scopus)

Abstract

Cellulose‐binding domains (CBDs) are structurally and functionally independent, noncatalytic modules found in many cellulose or hemicellulose degrading enzymes. Recent biotechnological applications of the CBDs include facilitated protein immobilization on cellulose supports.
In some occasions there have been concerns about the stability of the CBD driven immobilization. Here we have studied the chromatographic behavior of variants of the Trichoderma reesei cellobiohydrolase I CBD belonging to family I. Both CBDs fused to antibody fragments and isolated CBDs were studied and compared. Tritium labeling by reductive methylation was used as a sensitive detection method.
The fusion protein as well as the isolated CBD was found to leak from the column at a rate of 0.3–0.5% of the immobilized protein per column volume. However, the leakage could be overcome by using two CBDs instead of a single CBD for the immobilization. In this way leakage was reduced to less than 0.01% per column volume.
The improved immobilization could also be seen as a decreased migration of the protein down the column in extended washes.
Original languageEnglish
Pages (from-to)642-647
JournalBiotechnology and Bioengineering
Volume60
Issue number5
DOIs
Publication statusPublished - 1998
MoE publication typeA1 Journal article-refereed

Fingerprint

Cellulose
Immobilization
Fusion reactions
Proteins
Cellulose 1,4-beta-Cellobiosidase
Immobilized Proteins
Immunoglobulin Fragments
Methylation
Tritium
Labeling
Trichoderma
Antibodies
Enzymes
hemicellulose

Cite this

Linder, Markus ; Nevanen, Tarja ; Söderholm, Lotta ; Bengs, Outi ; Teeri, Tuula. / Improved immobilization of fusion proteins via cellulose-binding domains. In: Biotechnology and Bioengineering. 1998 ; Vol. 60, No. 5. pp. 642-647.
@article{08484532cc504d54a3382505f90feee4,
title = "Improved immobilization of fusion proteins via cellulose-binding domains",
abstract = "Cellulose‐binding domains (CBDs) are structurally and functionally independent, noncatalytic modules found in many cellulose or hemicellulose degrading enzymes. Recent biotechnological applications of the CBDs include facilitated protein immobilization on cellulose supports.In some occasions there have been concerns about the stability of the CBD driven immobilization. Here we have studied the chromatographic behavior of variants of the Trichoderma reesei cellobiohydrolase I CBD belonging to family I. Both CBDs fused to antibody fragments and isolated CBDs were studied and compared. Tritium labeling by reductive methylation was used as a sensitive detection method. The fusion protein as well as the isolated CBD was found to leak from the column at a rate of 0.3–0.5{\%} of the immobilized protein per column volume. However, the leakage could be overcome by using two CBDs instead of a single CBD for the immobilization. In this way leakage was reduced to less than 0.01{\%} per column volume. The improved immobilization could also be seen as a decreased migration of the protein down the column in extended washes.",
author = "Markus Linder and Tarja Nevanen and Lotta S{\"o}derholm and Outi Bengs and Tuula Teeri",
year = "1998",
doi = "10.1002/(SICI)1097-0290(19981205)60:5<642::AID-BIT15>3.0.CO;2-8",
language = "English",
volume = "60",
pages = "642--647",
journal = "Biotechnology and Bioengineering",
issn = "0006-3592",
publisher = "Wiley",
number = "5",

}

Improved immobilization of fusion proteins via cellulose-binding domains. / Linder, Markus (Corresponding Author); Nevanen, Tarja; Söderholm, Lotta; Bengs, Outi; Teeri, Tuula.

In: Biotechnology and Bioengineering, Vol. 60, No. 5, 1998, p. 642-647.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Improved immobilization of fusion proteins via cellulose-binding domains

AU - Linder, Markus

AU - Nevanen, Tarja

AU - Söderholm, Lotta

AU - Bengs, Outi

AU - Teeri, Tuula

PY - 1998

Y1 - 1998

N2 - Cellulose‐binding domains (CBDs) are structurally and functionally independent, noncatalytic modules found in many cellulose or hemicellulose degrading enzymes. Recent biotechnological applications of the CBDs include facilitated protein immobilization on cellulose supports.In some occasions there have been concerns about the stability of the CBD driven immobilization. Here we have studied the chromatographic behavior of variants of the Trichoderma reesei cellobiohydrolase I CBD belonging to family I. Both CBDs fused to antibody fragments and isolated CBDs were studied and compared. Tritium labeling by reductive methylation was used as a sensitive detection method. The fusion protein as well as the isolated CBD was found to leak from the column at a rate of 0.3–0.5% of the immobilized protein per column volume. However, the leakage could be overcome by using two CBDs instead of a single CBD for the immobilization. In this way leakage was reduced to less than 0.01% per column volume. The improved immobilization could also be seen as a decreased migration of the protein down the column in extended washes.

AB - Cellulose‐binding domains (CBDs) are structurally and functionally independent, noncatalytic modules found in many cellulose or hemicellulose degrading enzymes. Recent biotechnological applications of the CBDs include facilitated protein immobilization on cellulose supports.In some occasions there have been concerns about the stability of the CBD driven immobilization. Here we have studied the chromatographic behavior of variants of the Trichoderma reesei cellobiohydrolase I CBD belonging to family I. Both CBDs fused to antibody fragments and isolated CBDs were studied and compared. Tritium labeling by reductive methylation was used as a sensitive detection method. The fusion protein as well as the isolated CBD was found to leak from the column at a rate of 0.3–0.5% of the immobilized protein per column volume. However, the leakage could be overcome by using two CBDs instead of a single CBD for the immobilization. In this way leakage was reduced to less than 0.01% per column volume. The improved immobilization could also be seen as a decreased migration of the protein down the column in extended washes.

U2 - 10.1002/(SICI)1097-0290(19981205)60:5<642::AID-BIT15>3.0.CO;2-8

DO - 10.1002/(SICI)1097-0290(19981205)60:5<642::AID-BIT15>3.0.CO;2-8

M3 - Article

VL - 60

SP - 642

EP - 647

JO - Biotechnology and Bioengineering

JF - Biotechnology and Bioengineering

SN - 0006-3592

IS - 5

ER -