이매스교육 및 공지사항
Abstract
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.
1. 관련문헌 링크 : Development of an LC-MS/MS peptide mapping protocol for the
NISTmAb
2. 관련 tool 링크
: NIST Mass and Fragment Calculator v1.3
키워드 : peptide sequencing, NIST Mass
and Fragment Calculator v1.3