Date Approved
6-19-2024
Embargo Period
6-19-2024
Document Type
Thesis
Degree Name
Master of Science (M.S.) in Pharmaceutical Sciences
Department
Chemistry and Biochemistry
College
College of Science & Mathematics
Funder
AMT
Advisor
James Grinias, Ph.D.
Committee Member 1
Subash Jonnalagadda, Ph.D.
Committee Member 2
Gregory Caputo, Ph.D.
Keywords
Antibody; Glycosylation; HPLC; Peptides; Protein; Separation
Subject(s)
Liquid chromatography
Disciplines
Analytical Chemistry | Chemistry | Medicinal-Pharmaceutical Chemistry
Abstract
Using 1.5 mm inner diameter (i.d.) columns to bridge the gap between routinely used 2.1 mm i.d. columns and capillary bore columns allows for a sequential but significant increase in performance without the need for specialized equipment associated with very low dispersion liquid chromatography (LC) systems. These 1.5 mm i.d. columns balance an increase in sensitivity compared to 2.1 mm i.d. columns (theoretically doubling the time-domain peak area in mass sensitive detectors for the same mass load), while mitigating the efficiency losses due to extra-column dispersion effects that are commonly observed with 1.0 mm i.d. columns. Here, the use of 1.5 mm i.d. columns was applied to LC/UV analysis of small molecules and LC/MS analysis of recombinant monoclonal antibodies. The 1.5 mm i.d. column exhibited a two-to-threefold improvement in analyte peak area signal for small molecules as well as for intact, subunit, and peptide levels of antibody analysis. Still, the utility of capillary columns for the analysis of recombinant monoclonal antibodies and other high molecular weight molecules has its benefits. Improvements in column performance and ruggedness along with instrumentation improvement would increase the use of capillary systems. One modern trend in capillary instrumentation is miniaturization of the equipment to such a small size that it is portable and can be used inside or outside of a laboratory. Examples of separations using such an instrument are shown.
Recommended Citation
Libert, Benjamin, "IMPLEMENTING 1.5 MILLIMETER INTERNAL DIAMETER COLUMNS AND A PORTABLE CAPILLARY COLUMN INSTRUMENT INTO MONOCLONAL ANTIBODY ANALYTICAL WORKFLOWS" (2024). Theses and Dissertations. 3249.
https://rdw.rowan.edu/etd/3249
