Affinity enrichment; Breast cancer patient serum; Degradation study; LC-QTOF-MS; Therapeutic monoclonal antibody; Antibodies, Monoclonal; Peptides; Trastuzumab; Chromatography, Liquid/methods; Humans; Tandem Mass Spectrometry/methods; Breast Cancer; Cancer patients; In-vitro; In-vivo; LC-Q-ToF MS; Therapeutic monoclonal antibodies; Chromatography, Liquid; Tandem Mass Spectrometry; Analytical Chemistry; Environmental Chemistry; Biochemistry; Spectroscopy
Abstract :
[en] Degradation analysis of therapeutic mAb is of high interest for critical quality attributes assessment and biotransformation studies. However, some obstacles, including low in vivo concentrations of mAb and complex biological matrices containing IgGs, could seriously interfere with mAb bioanalysis. In this study, a bioanalytical platform was developed for studying in vitro/in vivo modifications of trastuzumab, in which specific capture on mimotope peptide modified material was combined with trypsin digestion and LC-QTOF-MS analysis. It is worth noting that this material exhibits high specificity, suitable dynamic binding capacity, very little non-specific protein adsorption, and thus provides good enrichment and quantification performances for trastuzumab from patient serums. In particular, this bioanalytical platform was successfully applied to the dynamic monitoring of modifications of trastuzumab, such as deamidation, isomerization, oxidation and cyclization. Except for the faster deamidation of LC-Asn-30 and HC-Asn-387/392/393 under serum incubation, similar degradation trends for other sites were observed in phosphate buffer and spiked serum. Differences of peptide modification degrees of trastuzumab in patient serums were also observed. The novel platform exhibited superior specificity than Protein A/G/L based analytical methods, lower cost and higher stability than antigen or anti-idiotypic antibody based analytical methods, ensuring the evaluation of modification sites.
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Lu, Li; Institute of Pharmaceutical Analysis, College of Pharmacy, Jinan University, Guangzhou, 510632, China, Laboratory for the Analysis of Medicines (LAM), Department of Pharmacy, CIRM, University of Liege, Avenue Hippocrate 15, B36 Tour 4 +3, 4000, Liège, Belgium
Liu, Xiao; Institute of Pharmaceutical Analysis, College of Pharmacy, Jinan University, Guangzhou, 510632, China
Zuo, Chengyi; Institute of Pharmaceutical Analysis, College of Pharmacy, Jinan University, Guangzhou, 510632, China
Zhou, Jingwei; Institute of Pharmaceutical Analysis, College of Pharmacy, Jinan University, Guangzhou, 510632, China
Zhu, Chendi; Institute of Pharmaceutical Analysis, College of Pharmacy, Jinan University, Guangzhou, 510632, China
Zhang, Zhang; Department of Pharmacy and Guangdong Province Key Laboratory of Pharmacodynamic Constituents of Traditional Chinese Medicine & New Drug Research, Jinan University, Guangzhou, 510632, China
Fillet, Marianne ; Université de Liège - ULiège > Département de pharmacie > Analyse des médicaments
Crommen, Jacques ; Université de Liège - ULiège > Département de pharmacie
Jiang, Zhengjin; Institute of Pharmaceutical Analysis, College of Pharmacy, Jinan University, Guangzhou, 510632, China, Department of Pharmacy and Guangdong Province Key Laboratory of Pharmacodynamic Constituents of Traditional Chinese Medicine & New Drug Research, Jinan University, Guangzhou, 510632, China. Electronic address: jzjjackson@hotmail.com
Wang, Qiqin; Institute of Pharmaceutical Analysis, College of Pharmacy, Jinan University, Guangzhou, 510632, China, Department of Pharmacy and Guangdong Province Key Laboratory of Pharmacodynamic Constituents of Traditional Chinese Medicine & New Drug Research, Jinan University, Guangzhou, 510632, China. Electronic address: qiqinxtu@163.com
Language :
English
Title :
In vitro/in vivo degradation analysis of trastuzumab by combining specific capture on HER2 mimotope peptide modified material and LC-QTOF-MS.
Guangzhou Science and Technology Program key projects
Funders :
NSCF - National Natural Science Foundation of China
Funding text :
This work was supported by the National Natural Science Foundation of China (Grant: 82173773 , 82073806 ), the Natural Science Foundation of Guangdong Province, China (Grant: 2021A0505030039 , 2022A1515011576 ), and Science and Technology Program of Guangzhou, China (Grant: 202102020729 ).
Kellie, J.F., Karlinsey, M.Z., Review of approaches and examples for monitoring biotransformation in protein and peptide therapeutics by MS. Bioanalysis 10 (2018), 1877–1890.
Schadt, S., Hauri, S., Lopes, F., Edelmann, M.R., Staack, R.F., Villaseñor, R., Kettenberger, H., Roth, A.B., Schuler, F., Richter, W.F., Funk, C., Are biotransformation studies of therapeutic proteins needed? Scientific considerations and technical challenges. Drug Metab. Dispos. 47 (2019), 1443–1456.
Nowak, C., Tiwari, A., Liu, H.C., Asparagine deamidation in a complementarity determining region of a recombinant monoclonal antibody in complex with antigen. Anal. Chem. 90 (2018), 6998–7003.
Wang, Y., Li, X., Liu, Y., Richardson, D., Li, H., Shameem, M., Yang, X., Simultaneous monitoring of oxidation, deamidation, isomerization, and glycosylation of monoclonal antibodies by liquid chromatography-mass spectrometry method with ultrafast tryptic digestion. mAbs 8 (2016), 1477–1486.
Olaleye, O., Spanov, B., Ford, R., Govorukhina, N., van de Merbel, N.C., Bischoff, R., Enrichment and liquid chromatography-mass spectrometry analysis of trastuzumab and pertuzumab using affimer reagents. Anal. Chem. 93 (2021), 13597–13605.
Yan, Y., Wei, H., Fu, Y., Jusuf, S., Zeng, M., Ludwig, R., Krystek, S.R. Jr., Chen, G.D., Tao, L., Das, T.K., Isomerization and oxidation in the complementarity-determining regions of a monoclonal antibody: a study of the modification-structure-function correlations by hydrogen-deuterium exchange mass spectrometry. Anal. Chem. 88 (2016), 2041–2050.
Mo, J., Yan, Q., Kwong So, C., Soden, T., Lewis, M.J., Hu, P., Understanding the impact of methionine oxidation on the biological functions of IgG1 antibodies using hydrogen/deuterium exchange mass spectrometry. Anal. Chem. 88 (2016), 9495–9502.
Bersin, L.M., Patel, S.M., Topp, E.M., Effect of ‘pH’ on the rate of pyroglutamate formation in solution and lyophilized solids. Mol. Pharm. 18 (2021), 3116–3124.
Yang, N., Tang, Q., Hu, P., Lewis, M.J., Use of in vitro systems to model in vivo degradation of therapeutic monoclonal antibodies. Anal. Chem. 90 (2018), 7896–7902.
Kaur, S., Bateman, K.P., Glick, J., Jairaj, M., Kellie, J.F., Sydor, J., Zeng, J.N., IQ consortium perspective: complementary LBA and LC-MS in protein therapeutics bioanalysis and biotransformation assessment. Bioanalysis 12 (2020), 257–270.
Li, Y., Monine, M., Huang, Y., Swann, P., Nestorov, I., Lyubarskaya, Y., Quantitation and pharmacokinetic modeling of therapeutic antibody quality attributes in human studies. mAbs 8 (2016), 1079–1087.
Zhang, Y., Wu, S., Li, Y., Comparative study of profiling post-translational modifications of a circulating antibody drug in human with different capture reagents. Biologicals 45 (2017), 93–95.
Bults, P., Bischoff, R., Bakker, H., Gietema, J.A., van de Merbel, N.C., LC-MS/MS-Based monitoring of in vivo protein biotransformation: quantitative determination of trastuzumab and its deamidation products in human plasma. Anal. Chem. 88 (2016), 1871–1877.
Ayyar, B.V., Arora, S., Murphy, C., O'Kennedy, R., Affinity chromatography as a tool for antibody purification. Methods 56 (2012), 116–129.
Lee, S.H., Hoshino, Y., Randall, A., Zeng, Z., Baldi, P., Doong, R.A., Shea, K.J., Engineered synthetic polymer nanoparticles as IgG affinity ligands. J. Am. Chem. Soc. 134 (2012), 15765–15772.
Liu, Y., Lu, Y., Liu, Z., Restricted access boronate affinity porous monolith as a protein A mimetic for the specific capture of immunoglobulin G. Chem. Sci. 3 (2012), 1467–1471.
Barroso, T., Hussain, A., Roque, A.C.A., Aguiar-Ricardo, A., Functional monolithic platforms: chromatographic tools for antibody purification. Biotechnol. J. 8 (2013), 671–681.
Wang, Q., Jin, H., Xia, D., Shao, H., Peng, K., Liu, X., Huang, H., Zhang, Q., Guo, J., Wang, Y., Crommen, J., Gan, N., Jiang, Z., Biomimetic polymer-based method for selective capture of C-reactive protein in biological fluids. ACS Appl. Mater. Interfaces 10 (2018), 41999–42008.
Yang, H., Gurgel, P.V., Carbonell, R.G., Purification of human immunoglobulin G via Fc-specific small peptide ligand affinity chromatography. J. Chromatogr. A 1216 (2009), 910–918.
Zhao, W.W., Shi, Q.H., Sun, Y., Dual-ligand affinity systems with octapeptide ligands for affinity chromatography of hIgG and monoclonal antibody. J. Chromatogr. A 1369 (2014), 64–72.
Xue, A., Zhao, W.W., Liu, X., Sun, Y., Affinity chromatography of human IgG with octapeptide ligands identified from eleven peptide-ligand candidates. Biochem. Eng. J. 107 (2016), 18–25.
Menegatti, S., Naik, A.D., Carbonell, R.G., The hidden potential of small synthetic molecules and peptides as affinity ligands for bioseparations. Pharm. Bioproc. 1 (2013), 467–485.
Yang, L., Zhang, L., Yan, L., Zheng, H., Lu, P., Chen, J., Dai, J., Sun, H., Xu, Y., Yang, T., Stability assessment of a new antithrombotic small peptide, Arg-Gly-Asp-Trp-Arg (RGDWR), and its derivative. Biotechnol. Lett. 39 (2017), 1183–1190.
Fang, Y.M., Lin, D.Q., Yao, S.J., Review on biomimetic affinity chromatography with short peptide ligands and its application to protein purification. J. Chromatogr. A 1571 (2018), 1–15.
He, Y., Xie, S., Yang, X., Yuan, R., Chai, Y., Electrochemical peptide biosensor based on in situ silver deposition for detection of prostate specific antigen. ACS Appl. Mater. Interfaces 7 (2015), 13360–13366.
Navarro-Sanchez, J., Argente-Garcia, A.I., Moliner-Martinez, Y., Roca-Sanjuan, D., Antypov, D., Campins-Falco, P., Rosseinsky, M.J., Marti-Gastaldo, C., Peptide metal-organic frameworks for enantioselective separation of chiral drugs. J. Am. Chem. Soc. 139 (2017), 4294–4297.
Du, K., Peptide immobilized monolith containing tentacle-type functionalized polymer chains for high-capacity binding of immunoglobulin G. J. Chromatogr. A 1374 (2014), 164–170.
Wang, X., Xia, D., Han, H., Peng, K., Zhu, P., Crommen, J., Wang, Q., Jiang, Z., Biomimetic small peptide functionalized affinity monoliths for monoclonal antibody purification. Anal. Chim. Acta 1017 (2018), 57–65.
Jiang, B., Liu, W., Qu, H., Meng, L., Song, S., Ouyang, T., Shou, C., A novel peptide isolated from a phage display peptide library with trastuzumab can mimic antigen epitope of HER-2. J. Biol. Chem. 280 (2005), 4656–4662.
Favoino, E., Prete, M., Catacchio, G., Conteduca, G., Perosa, F., CD20-Mimotope peptides: a model to define the molecular basis of epitope spreading. Int. J. Mol. Sci., 20, 2019, 1920.
Liu, W., Bennett, A.L., Ning, W., Tan, H., Berwanger, J.D., Zeng, X., Bruening, M.L., Monoclonal antibody capture and analysis using porous membranes containing immobilized peptide mimotopes. Anal. Chem. 90 (2018), 12161–12167.
Berwanger, J.D., Tan, H.Y., Jokhadze, G., Bruening, M.L., Determination of the serum concentrations of the monoclonal antibodies bevacizumab, rituximab, and panitumumab using porous membranes containing immobilized peptide mimotopes. Anal. Chem. 93 (2021), 7562–7570.
Chen, C.J., Tseng, M.C., Lin, H.J., Lin, T.W., Chen, Y.R., Visual indicator for surfactant abundance in MS-based membrane and general proteomics applications. Anal. Chem. 82 (2010), 8283–8290.
Spiga, O., Bernini, A., Scarselli, M., Giovannoni, L., Laschi, F., Neri, P., Bracci, L., Lozzi, L., Niccolai, N., Histidyl tags and structural stabilization of linear peptides. Spectrosc. Lett. 35 (2002), 111–118.
Majorek, K.A., Kuhn, M.L., Chruszcz, M., Anderson, W.F., Minor, W., Double trouble-Buffer selection and His-tag presence may be responsible for nonreproducibility of biomedical experiments. Protein Sci. 23 (2014), 1359–1368.
Wang, Q., Peng, K., Gan, N., Liu, X., Huang, H., Shao, H., Jin, H., Crommen, J., Jiang, Z., Rapid fabrication of versatile zwitterionic super-hydrophilic polymers by sole-monomer system for biomolecules separation. Chem. Eng. J., 396, 2020, 125121.
Luo, Q., Zou, H., Zhang, Q., Xiao, X., Ni, J., High-performance affinity chromatography with immobilization of protein A and L-histidine on molded monolith. Biotechnol. Bioeng. 80 (2002), 481–489.
Tehrani Najafian, F., Bibi, N.S., Islam, T., Fernandez-Lahore, M., A megaporous material harbouring a peptide ligand for affinity IgG purification. Electrophoresis 38 (2017), 2914–2921.
Fung, E.N., Bryan, P., Kozhich, A., Techniques for quantitative LC-MS/MS analysis of protein therapeutics: advances in enzyme digestion and immunocapture. Bioanalysis 8 (2016), 847–856.
Stoll, D.R., Harmes, D.C., Danforth, J., Wagner, E., Guillarme, D., Fekete, S., Beck, A., Direct identification of rituximab main isoforms and subunit analysis by online selective comprehensive two-dimensional liquid chromatography-mass spectrometry. Anal. Chem. 87 (2015), 8307–8315.
Chelius, D., Rehder, D.S., Bondarenko, P.V., Identification and characterization of deamidation sites in the conserved regions of human immunoglobulin gamma antibodies. Anal. Chem. 77 (2005), 6004–6011.
Yin, S., Pastuskovas, C.V., Khawli, L.A., Stults, J.T., Characterization of therapeutic monoclonal antibodies reveals differences between in vitro and in vivo time-course studies. Pharm. Res. (N. Y.) 30 (2013), 167–178.
Li, Y., Huang, Y., Ferrant, J., Lyubarskaya, Y., Zhang, Y., Li, S., Wu, S., Assessing in vivo dynamics of multiple quality attributes from a therapeutic IgG4 monoclonal antibody circulating in cynomolgus monkey. mAbs 8 (2016), 961–968.
Halvorsen, T.G., Reubsaet, L., Antibody based affinity capture LC-MS/MS in quantitative determination of proteins in biological matrices. TRAC Trend. Anal. Chem. 95 (2017), 132–139.
Huang, L., Lu, J., Wroblewski, V.J., Beals, J.M., Riggin, R.M., In vivo deamidation characterization of monoclonal antibody by LC/MS/MS. Anal. Chem. 77 (2005), 1432–1439.
