Evaluation of a Dual-Wavelength Size Exclusion HPLC Method With Improved Sensitivity to Detect Protein Aggregates and Its Use to Better Characterize Degradation Pathways of an IgG1 Monoclonal Antibody

2010 ◽  
Vol 99 (6) ◽  
pp. 2582-2597 ◽  
Author(s):  
Michael D. Bond ◽  
Mark E. Panek ◽  
Zheru Zhang ◽  
Dana Wang ◽  
Promod Mehndiratta ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Protein Aggregates ◽  
Hplc Method ◽  
Dual Wavelength ◽  
Size Exclusion ◽  
Degradation Pathways ◽  
Igg1 Monoclonal Antibody ◽  
Size Exclusion Hplc

A size-exclusion HPLC method for the determination of sodium chondroitin sulfate in pharmaceutical preparations

2003 ◽  
Vol 31 (6) ◽  
pp. 1229-1236 ◽  
Author(s):  
Don Woong Choi ◽  
Mi Jung Kim ◽  
Hee Sung Kim ◽  
Soo Hyun Chang ◽  
Gi Sook Jung ◽  
...  
Keyword(s):  
Chondroitin Sulfate ◽  
Pharmaceutical Preparations ◽  
Hplc Method ◽  
Size Exclusion ◽  
Size Exclusion Hplc

An improved size exclusion-HPLC method for molecular size distribution analysis of immunoglobulin G using sodium perchlorate in the eluent

2017 ◽  
Vol 138 ◽  
pp. 330-343 ◽  
Author(s):  
Hsiaoling Wang ◽  
Mark S. Levi ◽  
Alfred V. Del Grosso ◽  
William M. McCormick ◽  
Lokesh Bhattacharyya
Keyword(s):  
Size Distribution ◽  
Immunoglobulin G ◽  
Sodium Perchlorate ◽  
Molecular Size ◽  
Hplc Method ◽  
Size Exclusion ◽  
Distribution Analysis ◽  
Size Exclusion Hplc ◽  
Molecular Size Distribution

Size exclusion HPLC method for the determination of acidic fibroblast growth factor in viscous formulations

1997 ◽  
Vol 15 (12) ◽  
pp. 1929-1935 ◽  
Author(s):  
Mark W. Bruner ◽  
Joel Goldstein ◽  
C. Russell Middaugh ◽  
Marvin A. Brooks ◽  
David B. Volkin
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Hplc Method ◽  
Size Exclusion ◽  
Fibroblast Growth ◽  
Acidic Fibroblast Growth Factor ◽  
Size Exclusion Hplc

scholarly journals A Size Exclusion HPLC Method for Evaluating the Individual Impacts of Sugars and Organic Acids on Beverage Global Taste by Means of Calculated Dose-Over-Threshold Values

2014 ◽  
Vol 1 (3) ◽  
pp. 141-158 ◽  
Author(s):  
Luís Dias ◽  
Cédric Sequeira ◽  
Ana Veloso ◽  
Jorge Morais ◽  
Mara Sousa ◽  
...  
Keyword(s):  
Organic Acids ◽  
Hplc Method ◽  
Size Exclusion ◽  
Threshold Values ◽  
Calculated Dose ◽  
Size Exclusion Hplc ◽  
The Individual

scholarly journals Development of a Size-Exclusion HPLC Method with Evaporative Light-Scattering Detection for the Quantitation of Polysorbate 80 in Houttuynia cordata Injection

2010 ◽  
Vol 93 (3) ◽  
pp. 917-921 ◽  
Author(s):  
Yi Wu ◽  
Xiao D Li ◽  
Rui C Lin ◽  
Shao H Jin
Keyword(s):  
Light Scattering ◽  
Hplc Method ◽  
Size Exclusion ◽  
Polysorbate 80 ◽  
Good Precision ◽  
Evaporative Light Scattering Detector ◽  
Houttuynia Cordata ◽  
Light Scattering Detection ◽  
Evaporative Light Scattering ◽  
Size Exclusion Hplc

Abstract A rapid and accurate size-exclusion HPLC method for the quantitation of polysorbate 80 (PS80) in Houttuynia cordata injection, a Chinese traditional medicine, was developed and validated. The assay was conducted on an Agilent 1100 HPLC system with a TosoHaas TSKgel G2000 SWXL column (30 cm 7.8 mm, 5 m particle size) and an Alltech evaporative light-scattering detector (ELSD) 2000. The mobile phase was 20 mmoL/L ammonium acetateacetonitrile (90 + 10, v/v) delivered at a flow rate of 0.6 mL/min under isocratic conditions. The ELSD was operated in the impactor off mode, the drift tube temperature was set at 110C, and nitrogen flow was maintained at 2.3 L/min. The LOD was 0.25 mg/mL. Linearity was obtained between the log of concentration (C) and the log of peak area (Y) of PS80 in the range of 0.520 mg/mL according to the equation: Log Y = 1.4529 Log C 0.8232 (r2 = 0.9976). An RSD of 1.6 (n = 6) for the determination demonstrated the good precision of the optimized method. PS80 content in several commercial H. cordata injection products from different manufacturers was determined. The data for PS80 content is useful in evaluation of the safety of the products from different manufacturers.

scholarly journals Coelution of Other Proteins with Albumin during Size-Exclusion HPLC: Implications for Analysis of Urinary Albumin

2006 ◽  
Vol 52 (3) ◽  
pp. 389-397 ◽  
Author(s):  
Denis Sviridov ◽  
Bonnie Meilinger ◽  
Steven K Drake ◽  
Gerard T Hoehn ◽  
Glen L Hortin
Keyword(s):  
Mass Spectrometry ◽  
Reversed Phase ◽  
Hplc Method ◽  
Urinary Albumin ◽  
Size Exclusion ◽  
Albumin Concentration ◽  
Reversed Phase Hplc ◽  
Multiple Components ◽  
Size Exclusion Hplc ◽  
Phosphate Buffered Saline

Abstract Background: Size-exclusion HPLC has been used as an alternative to immunoassays for quantifying urinary albumin (microalbumin). Systematically higher values for the HPLC method have been proposed to result from nonimmunoreactive albumin. Methods: We evaluated separation of purified proteins and urinary components by size-exclusion HPLC using a Zorbax Bio Series GF-250 column eluted with phosphate-buffered saline. Urinary components eluting in the “albumin” peak were analyzed by mass spectrometry and reversed-phase HPLC. Results: Several proteins, such as transferrin, α1-proteinase inhibitor, α1-acid glycoprotein, and α2-HS glycoprotein, analyzed as purified components, were not resolved from albumin by size-exclusion HPLC. Peaks for other proteins, such as IgG and urinary components identified as dimers of α1-microglobulin and immunoglobulin light chains, overlapped with the albumin peak. Profiles of urine specimens showed variable amounts of components overlapping with albumin. Furthermore, the albumin peak obtained by size-exclusion HPLC was found by mass spectrometry and reversed-phase HPLC to contain multiple components in addition to albumin. Conclusions: Size-exclusion HPLC does not resolve albumin from several other proteins in urine. The albumin peak resolved by this technique, although predominantly composed of albumin, contains several coeluting globulins that would contribute to overestimation of albumin concentration by size-exclusion HPLC.

Evaluation of the Efficiency of Protein A Affinity Chromatography to Purify a Monoclonal Antibody for Cancer Treatment and its Purity Analysis

2020 ◽  
Vol 7 (2) ◽  
pp. 121-133
Author(s):  
Ayesha Akhtar ◽  
Shivakumar Arumugam ◽  
Shoaib Alam
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Affinity Chromatography ◽  
Protein A ◽  
Exchange Chromatography ◽  
Size Exclusion ◽  
High Yield ◽  
Staphylococcal Protein A ◽  
Purification Step ◽  
Purity Analysis

Background:: Protein A affinity chromatography is often employed as the most crucial purification step for monoclonal antibodies to achieve high yield with purity and throughput requirements. Introduction:: Protein A, also known as Staphylococcal protein A (SPA) is found in the cell wall of the bacteria staphylococcus aureus. It is one of the first discovered immunoglobulin binding molecules and has been extensively studied since the past few decades. The efficiency of Protein A affinity chromatography to purify a recombinant monoclonal antibody in a cell culture sample has been evaluated, which removes 99.0% of feed stream impurities. Materials and Method:: We have systematically evaluated the purification performance by using a battery of analytical methods SDS-PAGE (non-reduced and reduced sample), Cation Exchange Chromatography (CEX), Size-exclusion chromatography (SEC), and Reversed phased-Reduced Chromatography for a CHO-derived monoclonal antibody. Results and Discussion:: The analytical test was conducted to determine the impurity parameter, Host Cell Contaminating Proteins (HCP). It was evaluated to be 0.015ng/ml after the purification step; while initially, it was found to be 24.431ng/ml. Conclusion:: The tests showed a distinct decrease in the level of different impurities after the chromatography step. It can be concluded that Protein A chromatography is an efficient step in the purification of monoclonal antibodies.

Development of capillary size exclusion chromatography for the analysis of monoclonal antibody fragments extracted from human vitreous humor

2012 ◽  
Vol 1270 ◽  
pp. 111-117 ◽  
Author(s):  
Jennifer C. Rea ◽  
Yun Lou ◽  
Joel Cuzzi ◽  
Yuhua Hu ◽  
Isabella de Jong ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Size Exclusion Chromatography ◽  
Vitreous Humor ◽  
Antibody Fragments ◽  
Size Exclusion ◽  
Exclusion Chromatography ◽  
Capillary Size
Sign in / Sign up

Export Citation Format

Share Document