A simple procedure for preparing biotinylated immunoglobulin M from hybridoma culture medium.
Protein Expr Purif
; 225: 106595, 2025 Jan.
Article
em En
| MEDLINE
| ID: mdl-39197671
ABSTRACT
We previously reported a chromatography system for purifying immunoglobulin M (IgM) using N,N,N',N'-ethylenediaminetetrakis(methylenephosphonic acid)-modified zirconia particles that selectively absorb immunoglobulins. Here, we report a simple procedure for preparing biotinylated IgM from hybridoma culture medium using this zirconia-based chromatography system. The culture medium of an IgM-producing hybridoma cell line was used as the starting sample solution, and the IgM in the medium was concentrated and partially purified by zirconia chromatography. Next, 9-(biotinamido)-4,7-dioxanonanoic acid N-succinimidyl ester was added to react with the proteins in the sample. Subsequently, only the biotinylated IgM was isolated by Capto Core 400 polishing column chromatography. The entire process was easy to perform, could be completed within 2 h, and provided highly pure biotin-labeled IgM. This procedure is expected to be applicable to the labeling of IgM with various compounds and drugs.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Imunoglobulina M
/
Biotinilação
/
Meios de Cultura
/
Hibridomas
Limite:
Animals
Idioma:
En
Revista:
Protein Expr Purif
/
Protein expr. purif
/
Protein expression and purification
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
2025
Tipo de documento:
Article
País de publicação:
Estados Unidos