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Expression and Purification of Full-Length and Domain-Fragment Recombinant Pentraxin 3 (PTX3) Proteins from Mammalian and Bacterial Cells.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2020; Vol. 2132, pp. 65-74. - Publication Year :
- 2020
-
Abstract
- Although cell-based protein expression systems enable us a certain amount of protein suitable for subsequent biological experiments to be obtained, aggregates of the protein of interest are sometimes encountered during the purification procedure. Pentraxin 3 (PTX3), a member of the pentraxin family that is classified as a carbohydrate-binding protein based on its structure, comprises one of the humoral arms of the pattern recognition receptors that play an important role in the innate immune response. PTX3 comprises two domains; an N-terminal domain and a C-terminal domain. The C-terminal domain containing pentraxin signature has similar biological functions as other pentraxins such as C-reactive protein (CRP) and serum amyloid-P component (SAP). On the other side, the N-terminal domain is specific to PTX3. A supply of the PTX3 protein in full length or partial fragments is thus essential for the elucidation of its biological functions. Here we describe the expression and purification of recombinant PTX3. An arginine-containing buffer is essential for the elution of bacterially expressed PTX3 N-terminal domain to minimize aggregation. This method allows high-yield purification of full-length or domain-fragment recombinant PTX3 proteins for biological study.
- Subjects :
- Animals
Arginine chemistry
Buffers
C-Reactive Protein metabolism
CHO Cells
Cricetulus
Escherichia coli genetics
Humans
Protein Domains
Serum Amyloid P-Component metabolism
C-Reactive Protein genetics
Escherichia coli growth & development
Protein Engineering methods
Recombinant Proteins isolation & purification
Serum Amyloid P-Component genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 2132
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 32306315
- Full Text :
- https://doi.org/10.1007/978-1-0716-0430-4_7