NOTICE: Due to a lapse in annual appropriations, most of this website is not being updated. Learn more.
Form submissions will still be accepted but will not receive responses at this time. Sections of this site for programs using non-appropriated funds (such as NVLAP) or those that are excepted from the shutdown (such as CHIPS and NVD) will continue to be updated.
An official website of the United States government
Here’s how you know
Official websites use .gov
A .gov website belongs to an official government organization in the United States.
Secure .gov websites use HTTPS
A lock (
) or https:// means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites.
Catherine A. Mouchahoir, Mellisa Ly, Michaella Levy, Lisa E. Kilpatrick, Scott C. Lute, Karen W. Phinney, Lisa Marzilli, Kurt A. Brorson, Michael T. Boyne, Darryl Davis, John E. Schiel
Abstract
The primary sequence of a protein, including therapeutic monoclonal antibodies (mAbs), is a critical quality attribute that determines a great deal of its functionality and stability. Significant effort is devoted to determining the complete amino acid sequence of recombinant proteins of potential therapeutic benefit. Several methodologies may be employed to determine the fidelity of transcription and translation of the predicted protein. The focus of this chapter is to demonstrate primary structure confirmation using the recombinant human IgG1ҝ NIST mAb. Mass spectrometry based primary structure determination is covered in this chapter. It also will address the question of what necessary orthogonal characterization (additional enzymes, intact, and/or middle down) should be performed. A historical review of the methods employed to determine the sequence of proteins is included for completeness.
Citation
Current State-of-the-Art and Emerging Technologies for the Analysis of Monoclonal Antibodies
Mouchahoir, C.
, Ly, M.
, Levy, M.
, Kilpatrick, L.
, Lute, S.
, Phinney, K.
, Marzilli, L.
, Brorson, K.
, Boyne, M.
, Davis, D.
and Schiel, J.
(2015),
Determination of the Primary Sequence/ Structure, ACS Publications, Washington, DC, [online], https://doi.org/10.1021/bk-2015-1201.ch001
(Accessed October 1, 2025)