215810-47-2Relevant academic research and scientific papers
The use of turbulent flow chromatography for rapid, on-line analysis of tryptic digests
Couchman,Jones,Moniz
, p. 2140 - 2146 (2015/10/28)
Rationale Following digestion of proteins with trypsin, digests are typically subjected to further 'clean-up' prior to liquid chromatography/mass spectometry (LC/MS) analysis, in order to reduce the complexity of the digested matrix, as well as helping to remove residual denaturants and reduction/alkylation reagents prior to injection onto the analytical HPLC column. Often, this is carried out using off-line techniques, and is not ideally suited to high-throughput workloads, for example in clinical laboratories. Methods Bovine serum albumin (BSA) was used as a model protein. Following denaturation with urea, reduction/alkylation, and digestion with trypsin, the analytical recovery of a selection of proteotypic BSA peptides was assessed using a two-dimensional, turbulent flow chromatography method. Peptides were identified using a Q Exactive mass spectometer operating in full-scan mode. Results Total analysis time (including the on-line sample clean-up) was 15 min per injection. Aside from the most hydrophilic peptide selected, ATEEQLK, recovery using the turbulent flow chromatography systems was greater than 30% for all remaining peptides (N=17), and exceeded 50% for 12 of the 18 peptides studied. There was a broad correlation between the hydrophobicity factor and the observed recovery. Conclusions This study suggests that turbulent flow chromatography offers a rapid, on-line alternative to solid-phase extraction for the analysis of peptide digests by LC/MS. A wide range of column chemistries are available, and the technique can be further optimised for analyses which are targetted to specific peptides. As with turbulent flow chromatography for small-molecule workflows, this approach may be ideally suited to high-throughput applications, such as those which are emerging from within clinical laboratories.
A graphene-based multifunctional affinity probe for selective capture and sequential identification of different biomarkers from biosamples
Cheng, Gong,Wang, Zhi-Gang,Liu, Yan-Lin,Zhang, Ji-Lin,Sun, De-Hui,Ni, Jia-Zuan
supporting information, p. 10240 - 10242 (2012/11/13)
A novel multifunctional graphene-based affinity probe has been explored for selective capture of two different types of peptides from the biosamples for sequential detection. The Royal Society of Chemistry 2012.
