Protein extraction from human anagen head hairs 1-millimeter or less in total length

BioTechniques

Mehdi Moini¹  Joseph Donfack²  Traci L Carlson³  Brian A Eckenrode⁴  Brent M Allred⁵ 

[1] Education, Oak Ridge, TN 37830, USA;Forensic Science Research Unit, Federal Bureau of Investigation Laboratory Division, 2501 Investigation Parkway, Quantico, VA 22135, USA;;1Visiting Scientist Program, Oak Ridge Institute for Science &2Department of Forensic Sciences, George Washington University, 2100 Foxhall Road, NW Washington, District Of Columbia 20007, USA;;3Counterterrorism &

关键词: hair;    nano-flow liquid chromatography tandem-mass spectrometry;    nLC-MS/MS;    proteome;    proteomics;    protein extraction;   

DOI  :  10.2144/btn-2018-2004

来源: DOAJ

【 摘 要 】

A simple method for extracting protein from human anagen (i.e., actively growing hair stage) head hairs was developed in this study for cases of limited sample availability and/or studies of specific micro-features within a hair. The distinct feature segments of the hair from one donor were divided lengthwise (i.e., each of ∼200–400 μm) and then pooled for three individual hairs to form a total of eight composite hair samples (i.e., each of ∼1 mm or less in total length). The proteins were extracted, digested using trypsin, and characterized via nano-flow liquid chromatography tandem-mass spectrometry (nLC-MS/MS). A total of 63 proteins were identified from all eight protein samples analyzed of which 60% were keratin and keratin-associated proteins. The major hair keratins identified are consistent with previous studies using fluorescence in situ hybridization and nLC-MS/MS while requiring over 400–8000-fold less sample. The protein extraction method from micro-sized human head hairs described in this study will enable proteomic analysis of biological evidence for cases of limited sample availability and will complement hair research. For example, research seeking to develop alternative non-DNA based techniques for comparing questioned to known hairs, and understanding the biochemistry of hair decomposition.

【 授权许可】

Unknown