[소개]

본 논문은 바이오 의약품 단백질의 서열적인 특성을 규명하기 위하여 분석하는 접근 방법인 peptide mapping 분석법에 대하여 알기 쉽게 소개하는 논문입니다. 특히, HPLC-UV-MS 조합에서의 분석 조건에서 진행되는 내용이라 관련 분석법을 set-up 하시는 분들에게 도움이 되는 논문이라 소개해드립니다.

[초록]

Peptide mapping is a component of the analytical toolbox used within the biopharmaceutical industry to aid in the identity confirmation of a protein therapeutic and to monitor degradative events such as oxidation or deamidation. These methods offer the advantage of providing site-specific information regarding post-translational and chemical modifications that may arise during production, processing or storage. A number of such variations may also be induced by the sample preparation methods themselves which may confound the ability to accurately evaluate the true modification levels. One important focus when developing a peptide mapping method should therefore be the use of sample preparation conditions that will minimize the degree of artificial modifications induced. Unfortunately, the conditions that are amenable to effective reduction, alkylation and digestion are often the same conditions that promote unwanted modifications. Here we describe the optimization of a tryptic digestion protocol used for peptide mapping of the NISTmAb IgG1κ which addresses the challenge of balancing maximum digestion efficiency with minimum artificial modifications. The parameters on which we focused include buffer concentration, digestion time and temperature, as well as the source and type of trypsin (recombinant vs. pancreatic; bovine vs porcine) used. Using the optimized protocol we generated a peptide map of the NISTmAb which allowed us to confirm its identity at the level of primary structure.

논문링크 : Development of an LC-MS/MS peptide mapping protocol for the NISTmAb (nih.gov)

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<관련 분석항목>

Peptide mapping, Full amino acid sequencing, N-/C-terminal sequencing, Disulfide bond site determination, Free-thiol 분석, Protein/impurity protein ID, PTMs site determination

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