Validation of Peptide Mapping for Protein Identity and Genetic Stability. Biologics and Biotechnology Section, Pharmaceutical Research and Manufacturers of America

Allen, David P.; Baffi, R; Bausch, James; Bongers, Jacob; Costello, Maureen A.; Dougherty, John J.; Federici, Marcia; Garnick, Robert L.; Peterson, Sydney; Riggins, Ralph; Sewerin, K; Tuls, J

doi:10.1006/biol.1996.0034

Cited by 32 publications

(7 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This approach rePeptide mapping is frequently used to evaluate idenquired a low enzyme to protein ratio for the digestion to tity and/or purity of therapeutic recombinant proteins preclude enzyme-derived peaks produced by autolysis. (1). In this paper we describe the development of a However, the low enzyme concentrations necessitated tryptic mapping procedure for use with recombinant long incubation times at increased temperature.…”

Section: Generated Site Next To the Site Of Ligation This Mappingmentioning

confidence: 99%

See 1 more Smart Citation

Mapping of Recombinant Hemoglobin Using Immobilized Trypsin Cartridges

Lippincott¹,

Hess²,

Apostoł³

1997

Analytical Biochemistry

View full text Add to dashboard Cite

Section: Generated Site Next To the Site Of Ligation This Mappingmentioning

confidence: 99%

“…still maintaining quality. We examined a variety of approaches in order to obtain a reproducible peptide map which included various enzymes, alkylating agents, 1 To whom correspondence should be addressed.…”

Section: Generated Site Next To the Site Of Ligation This Mappingmentioning

confidence: 99%

Mapping of Recombinant Hemoglobin Using Immobilized Trypsin Cartridges

Lippincott¹,

Hess²,

Apostoł³

1997

Analytical Biochemistry

View full text Add to dashboard Cite

“…by comparison with an appropriate reference material that changes in the primary amino acid sequence have not occurred, confirming product consistency and/or genetic stability [23]. The method requires considerable expertise for performing peptide analysis in a quality control environment [23]. At present, only peptide maps generated by HPLC have been reported to be used for regulatory filings.…”

Section: Czementioning

confidence: 96%

“…Peptide maps are a regulatory requirement to demonstrate the identity of the protein and are also used to demonstrate genetic consistency in recombinant DNAderived proteins [23]. Peptide maps have also been used for determining changes in oxidation, deamidation, and the primary sequence of proteins by HPLC.…”

Section: Reviews Of Ce Used For the Analysis Of Recombinant Proteinsmentioning

confidence: 99%

Capillary Electrophoresis in the Development of Recombinant Protein Biopharmaceuticals

Chen¹,

Canova-Davis²

2001

CE in Biotechnology: Practical Applications for Protein and Peptide Analyses

View full text Add to dashboard Cite

The understanding of the physicochemical properties of proteins in solution is required to understand the separations taking place in the capillary. For example, during isoelectric focusing, it is important to ensure the lack of protein precipitation during fOCUSing and mobilization otherwise artifacts occur and the true resolution of charge variants is not achieved. Similarly, buffer conditions, including additives, composition of buffer, and pH can influence the resolution of product variants. With diligent efforts one can achieve desired separations. As knowledge of buffer conditions and use of these techniques increase, the methods will be improved.MECC can be useful for the characterization of recombinant proteins and monoclonal antibodies. However, the relatively long analyte migration times may be a deterrent for routine use in quality control situations. Pharmaceutical companies will employ new techniques which provide advantages over current technology. CE has advantages in certain applications. Applications which use slab gel techniques, IEF and SDS-PAGE can be replaced by the CE methods for formulation development, characterization of charge isoforms, and for purity determinations. CZE and clEF can be used for rapid identity tests which do not require peptide mapping. CZE has a tremendous advantage in analyzing basic proteins, and CZE and CE-SDS can be used to quantify product in the presence of interfering excipients. It is the responsibility of the analytical chemist to determine applications which are appropriate for CEo It is apparent that these are currently being defined for the biotechnology industry, and the usage of CE should continue to increase.

show abstract

“…Although analytical methodologies at both DNA and protein levels can be used to assess sequence variants, DNA sequencing methods have sensitivity limits of 10-15% and do not necessarily reflect the changes in the final product [17]. Historically, protein N-terminal sequencing by Edman degradation has been used on either electrophoretic gel bands or isolated chromatographic fractions to determine the sites and types of the variants [18][19][20][21]. More recently, mass spectrometry (MS) has become the predominant method for variant analysis [22,23], due to its high sensitivity, resolving power, and efficiency.…”

Section: Introductionmentioning

confidence: 99%