Label-free quantification of host cell protein impurity in recombinant hemoglobin materials.

ABSTRACT: Quantitative analysis relies on pure-substance primary calibrators with known mass fractions of impurity. Here, label-free quantification (LFQ) is being evaluated as a readily available, reliable method for determining the mass fraction of host cell proteins (HCPs) in bioengineered proteins which are intended for use as protein calibration standards. In this study a purified hemoglobin-A2 (HbA₂) protein, obtained through its overexpression in E. coli, was used. Two different materials were produced: natural and U¹⁵N-labeled HbA₂. For the quantification of impurities, precursor ion (MS1-) intensities were integrated over all E. coli proteins identified and divided by the intensities obtained for HbA₂. This ratio was calibrated against the correspo

SUBMITTER: Henrion A 

PROVIDER: S-EPMC10761545 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature