Quick purification of recombinant human truncated tau proteins for immunoanalysis

• Published in: Journal of immunological methods Vol. 185; no. 2; pp. 245 - 248
• Main Authors: Kontsekova, Eva, Cattaneo, Antonino, Novak, Michal
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 1995; Elsevier
• Subjects: Antibodies, Monoclonal - immunology; Biological and medical sciences; Enzyme-Linked Immunosorbent Assay - methods; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Hot Temperature; Human truncated τ protein; Humans; Molecular immunology; Molecular Weight; Peptide Fragments - immunology; Purification method; Recombinant Proteins - immunology; Recombinant Proteins - isolation & purification; tau Proteins - chemistry; tau Proteins - immunology; tau Proteins - isolation & purification; Techniques; Human truncated τ protein; Purification method; Human; Rapid technique; Heat treatment; Purification; Antigenicity; Tau protein; Truncated shape; Bacteria; Microtubule associated protein; Recombinant protein; Thermal stability
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/0022-1759(95)00120-Y

A simple and rapid purification method is described which exploits the heat stability of human tau (τ) protein to prepare truncated forms of this protein derived from bacteria. Bacterial cells expressing τ fragments were pelleted, resuspended in phosphate buffered saline and boiled for 5 min. After centrifugation the supernatant containing thermostable τ was filtered (0.45 μm) and used for immunoanalysis with monoclonal antibodies. The purified τ fragments exhibited identical antigenic properties as fragments isolated by a conventional procedure, based on ion exchange chromatography on phosphocellulose. In contrast to the conventional approach, our method is less complicated, cheaper and significantly reduces the time required for isolation of the recombinant τ fragments.