Isolating the whole complex of target proteins of FK506 using affinity resins from novel solid phases

• Published in: Analytical and bioanalytical chemistry Vol. 385; no. 1; pp. 122 - 127
• Main Authors: Takahashi, Teruki, Shiyama, Takaaki, Mori, Tomoko, Hosoya, Ken, Tanaka, Akito
• Format: Journal Article
• Language: English
• Published: Germany Berlin/Heidelberg : Springer-Verlag 01.05.2006; Springer Nature B.V
• Subjects: Affinity; Affinity resins; Animals; Bioactive compounds; Brain - metabolism; Calcineurin; Calcineurin - isolation & purification; Calcineurin - metabolism; calmodulin; Calmodulin - isolation & purification; Calmodulin - metabolism; Chromatography, Affinity - instrumentation; Chromatography, Affinity - methods; FK506; Molecular Structure; Poly(methacrylate); Proteins; Rats; Resins; Resins, Synthetic - chemistry; Solid phases; Substrate Specificity; Tacrolimus; Tacrolimus - metabolism; Tacrolimus Binding Protein 1A - isolation & purification; Tacrolimus Binding Protein 1A - metabolism; Tacrolimus Binding Proteins - isolation & purification; Tacrolimus Binding Proteins - metabolism
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-006-0335-3

The development of novel solid phases enabled us to create affinity resins that could be used to isolate the whole complex of target proteins responsible for the immunosuppressive effects of FK506 from rat brain lysate, whereas the affinity resins from commercially available matrices could not achieve this isolation. The results illustrate the enhanced effectiveness of the affinity resin made from this novel material at identifying the target protein of the bioactive compound compared to resins made from the well-known materials Affigel or Toyopearl. This effectiveness arises because the novel material is hydrophilic enough to reduce nonspecific binding proteins and because it has a higher density of ligands that capture the nonubiquitous target protein.