A universal surrogate peptide to enable LC-MS/MS bioanalysis of a diversity of human monoclonal antibody and human Fc-fusion protein drug candidates in pre-clinical animal studies

2012 ◽  
pp. n/a-n/a ◽  
Author(s):  
Michael T. Furlong ◽  
Zheng Ouyang ◽  
Steven Wu ◽  
James Tamura ◽  
Timothy Olah ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Fusion Protein ◽  
Animal Studies ◽  
Human Monoclonal Antibody ◽  
Protein Drug ◽  
Drug Candidates ◽  
Fc Fusion Protein ◽  
Surrogate Peptide

Dual universal peptide approach to bioanalysis of human monoclonal antibody protein drug candidates in animal studies

Bioanalysis ◽  
2013 ◽  
Vol 5 (11) ◽  
pp. 1363-1376 ◽  
Author(s):  
Michael T Furlong ◽  
Song Zhao ◽  
William Mylott ◽  
Rand Jenkins ◽  
Mian Gao ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Animal Studies ◽  
Human Monoclonal Antibody ◽  
Protein Drug ◽  
Drug Candidates

scholarly journals Comparison of Three Complementary Analytical Techniques for the Evaluation of the Biosimilar Comparability of a Monoclonal Antibody and an Fc-Fusion Protein

2021 ◽  
Vol 9 ◽  
Author(s):  
Alice Demelenne ◽  
Arij Ben Yahia ◽  
Delphine Lempereur ◽  
Jacques Crommen ◽  
Anne-Catherine Servais ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Fusion Protein ◽  
Method Development ◽  
Reference Product ◽  
Degradation Products ◽  
Stress Conditions ◽  
Size Exclusion ◽  
Mild Stress ◽  
Molecular Weight Determination ◽  
Fc Fusion Protein

In this work, a monoclonal antibody, adalimumab, and an Fc-fusion protein, etanercept, were studied and compared to one of their biosimilars. Samples submitted to stress conditions (agitation and high temperature) were used for method development. The developed methods were also applied to samples reduced by beta-mercaptoethanol to evaluate their capability to distinguish the expected species. Capillary gel electrophoresis (CGE), reversed-phase liquid chromatography (RPLC), and size-exclusion chromatography (SEC) methods coupled with UV detection were used to analyze the biopharmaceuticals. Their complementarity was investigated. For further molecular weight determination, SEC-multi angle light scattering and RPLC-quadrupole time-of-flight were occasionally used. For adalimumab, a larger amount of fragments and aggregates was observed in the biosimilar compared with the reference product. For etanercept, more related species were found in the reference product. Those three separation techniques showed good complementarity. Indeed, RPLC enabled the separation of hydrophilic and hydrophobic degradation products. CGE provided good selectivity for several adalimumab fragments, and SEC was useful for the analysis of aggregates and certain fragments that cannot be separated by the other approaches. Moreover, those formulations were submitted to mild stress conditions (30°C, 300 rpm for 4 h) that mimic shipping conditions. No additional peak was found under these conditions for the two studied biopharmaceuticals.

scholarly journals Bacterial Expression of a Human Monoclonal Antibody-Alkaline Phosphatase Conjugate Specific for Entamoeba histolytica

2004 ◽  
Vol 11 (1) ◽  
pp. 216-218 ◽  
Author(s):  
Hiroshi Tachibana ◽  
Masataka Takekoshi ◽  
Xun-Jia Cheng ◽  
Yuta Nakata ◽  
Tsutomu Takeuchi ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Alkaline Phosphatase ◽  
Fusion Protein ◽  
Entamoeba Histolytica ◽  
Human Monoclonal Antibody ◽  
Bacterial Expression ◽  
Human Antibody ◽  
Gene Encoding ◽  
E Coli ◽  
Fab Fragments

ABSTRACT We previously produced human monoclonal antibody Fab fragments specific to Entamoeba histolytica in Escherichia coli. In order to use these Fab fragments for diagnostic purposes, an expression vector to produce a fusion protein of Fab and alkaline phosphatase (PhoA) in E. coli was designed and constructed. The E. coli PhoA gene was fused to the 3′ terminus of the gene encoding the heavy-chain Fd region. The kappa and Fd genes from a previously prepared antibody clone, CP33, which is specific for the 260-kDa lectin of E. histolytica, were used as human antibody genes. When the fusion protein of CP33 and PhoA was incubated with paraformaldehyde-fixed trophozoites of E. histolytica and developed with a substrate, the trophozoites appeared to be stained. These results demonstrate the feasibility of bacterial expression of a human monoclonal antibody-PhoA conjugate specific for E. histolytica and that the antibody can be used to detect E. histolytica antigen without the use of chemically conjugated secondary antibodies.

P133 - Evaluating stability of human monoclonal antibody in cultured rat hepatocytes with a surrogate peptide appro

2020 ◽  
Vol 35 (1) ◽  
pp. S63
Author(s):  
Nadya Galeva ◽  
Reed Murbach ◽  
Krystal Gilligan ◽  
Kevin Westland ◽  
Seema Muranjan ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Human Monoclonal Antibody ◽  
Rat Hepatocytes ◽  
Cultured Rat Hepatocytes ◽  
Surrogate Peptide
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