Fast and high-throughput validation of protein products is of interest in biopharmaceutical industry. Top-down mass spectrometry (TDMS) allows for directly measuring of the intact form of proteins (i.e., proteoform) which have genetic mutations, alternative RNA splicing events and post-translational modifications. We’ve developed a new interface, called SampleStream, enabling fast and high-throughput measurement of proteoforms coupling to TDMS. The SampleStream interface consists of a fluidic channel that incorporates a molecular weight cutoff (MWCO) membrane to trap and concentrate proteins while allowing rapid buffer-exchange. The resulted proteins can be eluted from the channel then directly analyzed by high-resolution mass spectrometry. The platform offers the reduced sample processing time and quite similar sensitivity to conventional liquid chromatography (LC) technologies. We also combined immunoprecipitation with SampleStream-mass spectrometry (i.e., IP-SampleStream-MS) as a high-throughput workflow for the quantitative analysis of target proteins. We applied IP-SampleStream-MS to human serum samples to quantify proteoforms of apolipoproteins A-I (ApoA-I) and C-III (ApoC-III), which are associated with cardiometabolic charateristics such as high-density lipoprotein cholesterol (HDL-C) and obesity. We found proteoform-to-phenotype associations for ApoC-III, glycoproteoforms of which were characterized in this study.
저자
Hae-Min Park [ Department of Chemical Engineering and Applied Chemistry, Chungnam National University, Daejeon 34134, Republic of Korea ]
Corresponding Author
본 학회는 화학, 생화학, 분자생물학, 미생물학, 식품공학, 의학, 약학, 유전공학 및 생물공학, 환경 및 기타 공업 등 전 분야의 탄수화물관련 이론과 기술을 연구 발전시키고 산학협동을 통해 이를 보급하여 국내 관련 산업의 발전 및 국민생활의 과학화에 기여하고자 하며, 이러한 목표와 비젼의 실현을 위해 회원들이 적극적인 참여와 활동을 전개하고자 한다.