Large Molecule Bioanalysis Using Q-Tof Without Predigestion And Its Data Processing Challenges

Abstract

Selectivity and sensitivity were improved using different scan modes, such as TOF-MS and TOF-MS/MS.

Application of High-Resolution MS in The Quantification of A Therapeutic Monoclonal Antibody in Human Plasma

“…To simplify the process, Ramagiri et al highlighted three parameters that need to be optimized to achieve the best compromise between selectivity, specificity and sensitivity in the quantification by HRMS [17]. The parameters are: first, the XIC window size (2-50 mDa); second, selection of charge states, fragments and isotopic peaks to sum; and, third, comparison of sample (LLOQ) and matrix blank after varying the first two parameters.…”

“…The selectivity in this mode is determined by the narrow extraction mass window (XIC). This approach is most useful for quantification of large intact proteins due to its large scan windows or for quantification of peptides that show poor fragmentation and necessitates little method development and instrument optimization [17]. The second approach used in HRMS is the TOF-MS/MS mode, also known as MRM HR .…”

“…However, current advancements of high-resolution MS (HRMS) instrumentation, including enhanced scan speed, sensitivity and dynamic range have opened the door for LC-HRMS to play a significant role in the field of large-molecule quantitative bioanalysis [14][15][16][17][18]. Additionally, the increased mass range, characteristic of high-resolution time-of-flight (TOF) instruments, has been valuable in the bioanalysis of intact proteins of higher molecular weight [16].…”

Quantitation of Human Peptides and Proteins Via MS: Review of Analytically Validated Assays

“…future science group Quantitation of human peptides & proteins via MS: review of analytically validated assays Review an opportunity for full scan assays that can sum ion intensities from multiple charge states [36]. Peptide quantitation using LC-MS is rapidly evolving, and the number of circulating peptides quantitated with this technique continues to grow rapidly.…”

Application and Challenges in Using LC–MS Assays for Absolute Quantitative Analysis of Therapeutic Proteins in Drug Discovery

“…Compared with small molecules, proteins present additional bioanalytical challenges due to the complexity of the matrix resulting from the post-translational modifications and proteolytic cleavage of the therapeutic protein by peptidases. HRMS offers valuable features for resolving some of these bioanalytical challenges [86,87].…”