Chromatographic analysis of the acidic and basic species of recombinant monoclonal antibodies

mAbs

Volumn 4, Issue 5, 2012, Pages 578-585

  Du, Yi ^a   Walsh, Alison ^a   Ehrick, Robin ^a   Xu, Wei ^a   May, Kimberly ^a   Liu, Hongcheng ^a  

^a MERCK RESEARCH LABORATORIES   (United States)

Author keywords

Acidic species; Basic species; Posttranslational modifications; Recombinant monoclonal antibody

Indexed keywords

IMMUNOGLOBULIN G1 ANTIBODY; IMMUNOGLOBULIN G4; LYSINE; MANNOSE; MONOCLONAL ANTIBODY; RECOMBINANT ANTIBODY; SIALIC ACID;

ACIDITY; ALKALINITY; AMINO ACID SEQUENCE; ANTIBODY STRUCTURE; BINDING AFFINITY; CARBOXY TERMINAL SEQUENCE; CHEMICAL MODIFICATION; CHROMATOGRAPHY; DEAMINATION; DIFFERENTIAL SCANNING CALORIMETRY; DISULFIDE BOND; GLYCATION; GLYCOSYLATION; HEAVY CHAIN; HUMAN; LIGHT CHAIN; MOLECULAR WEIGHT; NONHUMAN; PROTEIN PROCESSING; RECEPTOR BINDING; REVIEW;

AMINO ACID SEQUENCE; ANIMALS; ANTIBODIES, MONOCLONAL; CHROMATOGRAPHY, ION EXCHANGE; HUMANS; HYDROGEN-ION CONCENTRATION; MICE; PROTEIN PROCESSING, POST-TRANSLATIONAL; RECOMBINANT PROTEINS;

EID: 84866526223     PISSN: 19420862     EISSN: 19420870     Source Type: Journal    
DOI: 10.4161/mabs.21328     Document Type: Review

References (70)

1. Perkins M, Theiler R, Lunte S, Jeschke M. Determination of the origin of charge heterogeneity in a murine monoclonal antibody. Pharm Res 2000; 17:1110-7; PMID:11087044; http://dx.doi.org/10.1023/A:1026461830617.
2. Harris RJ, Kabakoff B, Macchi FD, Shen FJ, Kwong M, Andya JD, et al. Identification of multiple sources of charge heterogeneity in a recombinant antibody. J Chromatogr B Biomed Sci Appl 2001; 752:233-45; PMID:11270864; http://dx.doi.org/10.1016/S0378-4347(00)00548-X.
3. Moorhouse KG, Nashabeh W, Deveney J, Bjork NS, Mulkerrin MG, Ryskamp T. Validation of an HPLC method for the analysis of the charge heterogeneity of the recombinant monoclonal antibody IDECC2B8 after papain digestion. J Pharm Biomed Anal 1997; 16:593-603; PMID:9502155; http://dx.doi.org/10.1016/S0731-7085(97) 00178-7.
4. Triguero D, Buciak JB, Yang J, Pardridge WM. Bloodbrain barrier transport of cationized immunoglobulin G: enhanced delivery compared to native protein. Proc Natl Acad Sci U S A 1989; 86:4761-5; PMID:2734318; http://dx.doi.org/10. 1073/pnas.86.12.4761.
5. Pardridge WM, Bickel U, Buciak J, Yang J, Diagne A, Aepinus C. Cationization of a monoclonal antibody to the human immunodeficiency virus REV protein enhances cellular uptake but does not impair antigen binding of the antibody. Immunol Lett 1994; 42:191-5; PMID:7890319; http://dx.doi.org/10.1016/ 0165-2478(94)90085-X.
6. Pardridge WM, Kang YS, Diagne A, Zack JA. Cationized hyperimmune immunoglobulins: pharmacokinetics, toxicity evaluation and treatment of human immunodeficiency virus-infected human-peripheral blood lymphocytes-severe combined immune deficiency mice. J Pharmacol Exp Ther 1996; 276:246-52; PMID:8558438.
7. Hong G, Bazin-Redureau MI, Scherrmann JM. Pharmacokinetics and organ distribution of cationized colchicine-specific IgG and Fab fragments in rat. J Pharm Sci 1999; 88:147-53; PMID:9874717; http://dx.doi.org/10.1021/js970335n.
8. Rodwell JD, Alvarez VL, Lee C, Lopes AD, Goers JW, King HD, et al. Site-specific covalent modification of monoclonal antibodies: in vitro and in vivo evaluations. Proc Natl Acad Sci U S A 1986; 83:2632-6; PMID:3458222; http://dx.doi.org/10.1073/pnas.83.8.2632.
9. Gangopadhyay A, Petrick AT, Thomas P. Modification of antibody isoelectric point affects biodistribution of 111-indium-labeled antibody. Nucl Med Biol 1996; 23:257-61; PMID:8782234; http://dx.doi.org/10.1016/0969-8051(95) 02057-8.
10. Khawli LA, Mizokami MM, Sharifi J, Hu P, Epstein AL. Pharmacokinetic characteristics and biodistribution of radioiodinated chimeric TNT-1, -2, and -3 monoclonal antibodies after chemical modification with biotin. Cancer Biother Radiopharm 2002; 17:359-70; PMID:12396700; http://dx.doi.org/10.1089/ 108497802760363150.
11. Lee HJ, Pardridge WM. Monoclonal antibody radiopharmaceuticals: cationization, pegylation, radiometal chelation, pharmacokinetics, and tumor imaging. Bioconjug Chem 2003; 14:546-53; PMID:12757378; http://dx.doi.org/10. 1021/bc0256648.
12. Perera RM, Zoncu R, Johns TG, Pypaert M, Lee FT, Mellman I, et al. Internalization, intracellular trafficking, and biodistribution of monoclonal antibody 806: a novel anti-epidermal growth factor receptor antibody. Neoplasia 2007; 9:1099-110; PMID:18084617; http://dx.doi.org/10.1593/neo.07721.
13. Khawli LA, Glasky MS, Alauddin MM, Epstein AL. Improved tumor localization and radioimaging with chemically modified monoclonal antibodies. Cancer Biother Radiopharm 1996; 11:203-15; PMID:10851539; http://dx.doi.org/10. 1089/cbr.1996.11.203.
14. Khawli LA, Goswami S, Hutchinson R, Kwong ZW, Yang J, Wang X, et al. Charge variants in IgG1: Isolation, characterization, in vitro binding properties and pharmacokinetics in rats. MAbs 2010; 2:613-24; PMID:20818176; http://dx.doi.org/10.4161/mabs.2.6.13333.
15. Vlasak J, Ionescu R. Heterogeneity of monoclonal antibodies revealed by charge-sensitive methods. Curr Pharm Biotechnol 2008; 9:468-81; PMID:19075686; http://dx.doi.org/10.2174/138920108786786402.
16. Liu H, Gaza-Bulseco G, Faldu D, Chumsae C, Sun J. Heterogeneity of monoclonal antibodies. J Pharm Sci 2008; 97:2426-47; PMID:17828757; http://dx.doi.org/10.1002/jps.21180.
17. Lyubarskaya Y, Houde D, Woodard J, Murphy D, Mhatre R. Analysis of recombinant monoclonal antibody isoforms by electrospray ionization mass spectrometry as a strategy for streamlining characterization of recombinant monoclonal antibody charge heterogeneity. Anal Biochem 2006; 348:24-39; PMID:16289440; http://dx.doi.org/10.1016/j. ab.2005.10.003.
18. Beck A, Bussat MC, Zorn N, Robillard V, Klinguer-Hamour C, Chenu S, et al. Characterization by liquid chromatography combined with mass spectrometry of monoclonal anti-IGF-1 receptor antibodies produced in CHO and NS0 cells. J Chromatogr B Analyt Technol Biomed Life Sci 2005; 819:203-18; PMID:15833284; http://dx.doi.org/10.1016/j.jchromb.2004.06.052.
19. Harris RJ. Processing of C-terminal lysine and arginine residues of proteins isolated from mammalian cell culture. J Chromatogr A 1995; 705:129-34; PMID:7620566; http://dx.doi.org/10.1016/0021-9673(94)01255-D.
20. Santora LC, Krull IS, Grant K. Characterization of recombinant human monoclonal tissue necrosis factoralpha antibody using cation-exchange HPLC and capillary isoelectric focusing. Anal Biochem 1999; 275:98-108; PMID:10542114; http://dx.doi.org/10.1006/ abio.1999.4275.
21. Santora LC, Stanley K, Krull IS, Grant K. Characterization of maleuric acid derivatives on transgenic human monoclonal antibody due to post-secretional modifications in goat milk. Biomed Chromatogr 2006; 20:843-56; PMID:16425344; http://dx.doi.org/10.1002/bmc.603.
22. Antes B, Amon S, Rizzi A, Wiederkum S, Kainer M, Szolar O, et al. Analysis of lysine clipping of a humanized Lewis-Y specific IgG antibody and its relation to Fc-mediated effector function. J Chromatogr B Analyt Technol Biomed Life Sci 2007; 852:250-6; PMID:17296336; http://dx.doi.org/10.1016/j. jchromb.2007.01.024.
23. Yan B, Steen S, Hambly D, Valliere-Douglass J, Vanden Bos T, Smallwood S, et al. Succinimide formation at Asn 55 in the complementarity determining region of a recombinant monoclonal antibody IgG1 heavy chain. J Pharm Sci 2009; 98:3509-21; PMID:19475547; http://dx.doi.org/10.1002/jps.21655.
24. Weitzhandler M, Farnan D, Horvath J, Rohrer JS, Slingsby RW, Avdalovic N, et al. Protein variant separations by cation-exchange chromatography on tentacle-type polymeric stationary phases. J Chromatogr A 1998; 828:365-72; PMID:9916317; http://dx.doi.org/10.1016/S0021-9673(98)00521-4.
25. Zhang T, Bourret J, Cano T. Isolation and characterization of therapeutic antibody charge variants using cation exchange displacement chromatography. J Chromatogr A 2011; 1218:5079-86; PMID:21700290; http://dx.doi.org/10.1016/j. chroma.2011.05.061.
26. Alvarez M, Tremintin G, Wang J, Eng M, Kao YH, Jeong J, et al. On-line characterization of monoclonal antibody variants by liquid chromatography-mass spectrometry operating in a two-dimensional format. Anal Biochem 2011; 419:17-25; PMID:21867674; http://dx.doi.org/10.1016/j.ab.2011.07.033.
27. Vlasak J, Bussat MC, Wang S, Wagner-Rousset E, Schaefer M, Klinguer-Hamour C, et al. Identification and characterization of asparagine deamidation in the light chain CDR1 of a humanized IgG1 antibody. Anal Biochem 2009; 392:145-54; PMID:19497295; http://dx.doi.org/10.1016/j.ab.2009.05.043.
28. Huang L, Lu J, Wroblewski VJ, Beals JM, Riggin RM. In vivo deamidation characterization of monoclonal antibody by LC/MS/MS. Anal Chem 2005; 77:1432-9; PMID:15732928; http://dx.doi.org/10.1021/ ac0494174.
29. Zheng JY, Janis LJ. Influence of pH, buffer species, and storage temperature on physicochemical stability of a humanized monoclonal antibody LA298. Int J Pharm 2006; 308:46-51; PMID:16316730; http://dx.doi.org/10.1016/j. ijpharm.2005.10.024.
30. Kim J, Jones L, Taylor L, Kannan G, Jackson F, Lau H, et al. Characterization of a unique IgG1 mAb CEX profile by limited Lys-C proteolysis/CEX separation coupled with mass spectrometry and structural analysis. J Chromatogr B Analyt Technol Biomed Life Sci 2010; 878:1973-81; PMID:20554483; http://dx.doi. org/10.1016/j.jchromb.2010.05.032.
31. Harris R, Shire SJ, Winter C. Commercial manufacturing scale formulation and analytical characterization of therapeutic recombinant antibodies. Drug Dev Res 2004; 61:137-54; http://dx.doi.org/10.1002/ddr.10344.
32. Chelius D, Rehder DS, Bondarenko PV. Identification and characterization of deamidation sites in the conserved regions of human immunoglobulin gamma antibodies. Anal Chem 2005; 77:6004-11; PMID:16159134; http://dx.doi.org/10. 1021/ac050672d.
33. Terashima I, Koga A, Nagai H. Identification of deamidation and isomerization sites on pharmaceutical recombinant antibody using H(2)(18)O. Anal Biochem 2007; 368:49-60; PMID:17617368; http://dx.doi.org/10.1016/j.ab.2007.05. 012.
34. Gaza-Bulseco G, Li B, Bulseco A, Liu HC. Method to differentiate asn deamidation that occurred prior to and during sample preparation of a monoclonal antibody. Anal Chem 2008; 80:9491-8; PMID:19072263; http://dx.doi.org/10.1021/ ac801617u.
35. Sinha S, Zhang L, Duan S, Williams TD, Vlasak J, Ionescu R, et al. Effect of protein structure on deamidation rate in the Fc fragment of an IgG1 monoclonal antibody. Protein Sci 2009; 18:1573-84; PMID:19544580; http://dx.doi.org/10.1002/pro.173.
36. Gandhi S, Ren D, Xiao G, Bondarenko P, Sloey C, Ricci MS, et al. Elucidation of degradants in acidic peak of cation exchange chromatography in an IgG1 monoclonal antibody formed on long-term storage in a liquid formulation. Pharm Res 2012; 29:209-24; PMID:21845507; http://dx.doi.org/10.1007/s11095-011- 0536-0.
37. Wang L, Amphlett G, Lambert JM, Blättler W, Zhang W. Structural characterization of a recombinant monoclonal antibody by electrospray time-of-flight mass spectrometry. Pharm Res 2005; 22:1338-49; PMID:16078144; http://dx.doi.org/10.1007/s11095-005-5267-7.
38. Mukherjee R, Adhikary L, Khedkar A, Iyer H. Probing deamidation in therapeutic immunoglobulin gamma (IgG1) by 'bottom-up' mass spectrometry with electron transfer dissociation. Rapid Commun Mass Spectrom 2010; 24:879-84; PMID:20196189; http://dx.doi.org/10.1002/rcm.4464.
39. Zhang W, Czupryn MJ. Analysis of isoaspartate in a recombinant monoclonal antibody and its charge isoforms. J Pharm Biomed Anal 2003; 30:1479-90; PMID:12467919; http://dx.doi.org/10.1016/S0731-7085(02)00479-X.
40. Wypych J, Li M, Guo A, Zhang Z, Martinez T, Allen MJ, et al. Human IgG2 antibodies display disulfide-mediated structural isoforms. J Biol Chem 2008; 283:16194-205; PMID:18339624; http://dx.doi.org/10.1074/jbc.M709987200.
41. Martinez T, Guo A, Allen MJ, Han M, Pace D, Jones J, et al. Disulfide connectivity of human immunoglobulin G2 structural isoforms. Biochemistry 2008; 47:7496-508; PMID:18549248; http://dx.doi.org/10.1021/ bi800576c.
42. Pristatsky P, Cohen SL, Krantz D, Acevedo J, Ionescu R, Vlasak J. Evidence for trisulfide bonds in a recombinant variant of a human IgG2 monoclonal antibody. Anal Chem 2009; 81:6148-55; PMID:19591437; http://dx.doi.org/10.1021/ac9006254.
43. Lam XM, Yang JY, Cleland JL. Antioxidants for prevention of methionine oxidation in recombinant monoclonal antibody HER2. J Pharm Sci 1997; 86:1250-5; PMID:9383735; http://dx.doi.org/10.1021/js970143s.
44. Banks DD, Hambly DM, Scavezze JL, Siska CC, Stackhouse NL, Gadgil HS. The effect of sucrose hydrolysis on the stability of protein therapeutics during accelerated formulation studies. J Pharm Sci 2009; 98:4501-10; PMID:19388069; http://dx.doi.org/10.1002/jps.21749.
45. Miller AK, Hambly DM, Kerwin BA, Treuheit MJ, Gadgil HS. Characterization of site-specific glycation during process development of a human therapeutic monoclonal antibody. J Pharm Sci 2011; 100:2543-50; PMID:21287557; http://dx.doi.org/10.1002/ jps.22504.
46. Quan C, Alcala E, Petkovska I, Matthews D, Canova-Davis E, Taticek R, et al. A study in glycation of a therapeutic recombinant humanized monoclonal antibody: where it is, how it got there, and how it affects charge-based behavior. Anal Biochem 2008; 373:179-91; PMID:18158144; http://dx.doi.org/10. 1016/j. ab.2007.09.027.
47. Lau H, Pace D, Yan B, McGrath T, Smallwood S, Patel K, et al. Investigation of degradation processes in IgG1 monoclonal antibodies by limited proteolysis coupled with weak cation-exchange HPLC. J Chromatogr B Analyt Technol Biomed Life Sci 2010; 878:868-76; PMID:20206584; http://dx.doi.org/10. 1016/j. jchromb.2010.02.003.
48. Banks DD, Gadgil HS, Pipes GD, Bondarenko PV, Hobbs V, Scavezze JL, et al. Removal of cysteinylation from an unpaired sulfhydryl in the variable region of a recombinant monoclonal IgG1 antibody improves homogeneity, stability, and biological activity. J Pharm Sci 2008; 97:775-90; PMID:17786988; http://dx.doi.org/10.1002/jps.21014.
49. Dillon TM, Ricci MS, Vezina C, Flynn GC, Liu YD, Rehder DS, et al. Structural and functional characterization of disulfide isoforms of the human IgG2 subclass. J Biol Chem 2008; 283:16206-15; PMID:18339626; http://dx.doi.org/10.1074/jbc.M709988200.
50. Dick LW Jr., Qiu D, Mahon D, Adamo M, Cheng KC. C-terminal lysine variants in fully human monoclonal antibodies: investigation of test methods and possible causes. Biotechnol Bioeng 2008; 100:1132-43; PMID:18553400; http://dx.doi.org/10.1002/bit.21855.
51. Ouellette D, Alessandri L, Chin A, Grinnell C, Tarcsa E, Radziejewski C, et al. Studies in serum support rapid formation of disulfide bond between unpaired cysteine residues in the VH domain of an immunoglobulin G1 molecule. Anal Biochem 2010; 397:37- 47; PMID:19766583; http://dx.doi.org/10.1016/j.ab. 2009.09.027.
52. Wakankar AA, Borchardt RT, Eigenbrot C, Shia S, Wang YJ, Shire SJ, et al. Aspartate isomerization in the complementarity-determining regions of two closely related monoclonal antibodies. Biochemistry 2007; 46:1534-44; PMID:17279618; http://dx.doi.org/10.1021/bi061500t.
53. Cacia J, Keck R, Presta LG, Frenz J. Isomerization of an aspartic acid residue in the complementarity-determining regions of a recombinant antibody to human IgE: identification and effect on binding affinity. Biochemistry 1996; 35:1897-903; PMID:8639672; http://dx.doi.org/10.1021/bi951526c.
54. Huang HZ, Nichols A, Liu D. Direct identification and quantification of aspartyl succinimide in an IgG2 mAb by RapiGest assisted digestion. Anal Chem 2009; 81:1686-92; PMID:19146457; http://dx.doi.org/10.1021/ac802708s.
55. Xiao G, Bondarenko PV, Jacob J, Chu GC, Chelius D. 18O labeling method for identification and quantification of succinimide in proteins. Anal Chem 2007; 79:2714-21; PMID:17313184; http://dx.doi.org/10.1021/ac0617870.
56. Chu GC, Chelius D, Xiao G, Khor HK, Coulibaly S, Bondarenko PV. Accumulation of succinimide in a recombinant monoclonal antibody in mildly acidic buffers under elevated temperatures. Pharm Res 2007; 24:1145-56; PMID:17385019; http://dx.doi.org/10.1007/s11095-007-9241-4.
57. Sreedhara A, Cordoba A, Zhu Q, Kwong J, Liu J. Characterization of the isomerization products of aspartate residues at two different sites in a monoclonal antibody. Pharm Res 2012; 29:187-97; PMID:21809161; http://dx.doi.org/10.1007/s11095-011-0534-2.
58. Chumsae C, Gaza-Bulseco G, Sun J, Liu H. Comparison of methionine oxidation in thermal stability and chemically stressed samples of a fully human monoclonal antibody. J Chromatogr B Analyt Technol Biomed Life Sci 2007; 850:285-94; PMID:17182291; http://dx.doi.org/10.1016/j.jchromb.2006.11.050.
59. Teshima G, Li MX, Danishmand R, Obi C, To R, Huang C, et al. Separation of oxidized variants of a monoclonal antibody by anion-exchange. J Chromatogr A 2011; 1218:2091-7; PMID:21145555; http://dx.doi.org/10.1016/j.chroma.2010.10. 107.
60. Johnson KA, Paisley-Flango K, Tangarone BS, Porter TJ, Rouse JC. Cation exchange-HPLC and mass spectrometry reveal C-terminal amidation of an IgG1 heavy chain. Anal Biochem 2007; 360:75- 83; PMID:17113563; http://dx.doi.org/10.1016/ j.ab.2006.10.012.
61. Kaschak T, Boyd D, Lu F, Derfus G, Kluck B, Nogal B, et al. Characterization of the basic charge variants of a human IgG1: effect of copper concentration in cell culture media. MAbs 2011; 3:577-83; PMID:22123059; http://dx.doi.org/10.4161/mabs.3.6.17959.
62. Ren D, Zhang J, Pritchett R, Liu H, Kyauk J, Luo J, et al. Detection and identification of a serine to arginine sequence variant in a therapeutic monoclonal antibody. J Chromatogr B Analyt Technol Biomed Life Sci 2011; 879:2877-84; PMID:21900054; http://dx.doi.org/10.1016/j.jchromb.2011.08.015.
63. Gaza-Bulseco G, Bulseco A, Chumsae C, Liu H. Characterization of the glycosylation state of a recombinant monoclonal antibody using weak cation exchange chromatography and mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci 2008; 862:155-60; PMID:18164669; http://dx.doi.org/10.1016/j. jchromb.2007.12.001.
64. Liu H, Bulseco GG, Sun J. Effect of posttranslational modifications on the thermal stability of a recombinant monoclonal antibody. Immunol Lett 2006; 106:144- 53; PMID:16831470; http://dx.doi.org/10.1016/j.imlet.2006.05.011.
65. Harris RJ. Heterogeneity of recombinant antibodies: linking structure to function. Dev Biol (Basel) 2005; 122:117-27; PMID:16375256.
66. Liu D, Ren D, Huang H, Dankberg J, Rosenfeld R, Cocco MJ, et al. Structure and stability changes of human IgG1 Fc as a consequence of methionine oxidation. Biochemistry 2008; 47:5088-100; PMID:18407665; http://dx.doi.org/10. 1021/bi702238b.
67. Bertolotti-Ciarlet A, Wang W, Lownes R, Pristatsky P, Fang Y, McKelvey T, et al. Impact of methionine oxidation on the binding of human IgG1 to Fc Rn and Fc gamma receptors. Mol Immunol 2009; 46:1878-82; PMID:19269032; http://dx.doi.org/10.1016/j.molimm.2009.02.002.
68. Pan H, Chen K, Chu L, Kinderman F, Apostol I, Huang G. Methionine oxidation in human IgG2 Fc decreases binding affinities to protein A and FcRn. Protein Sci 2009; 18:424-33; PMID:19165723; http:// dx.doi.org/10.1002/pro.45.
69. Gaza-Bulseco G, Faldu S, Hurkmans K, Chumsae C, Liu H. Effect of methionine oxidation of a recombinant monoclonal antibody on the binding affinity to protein A and protein G. J Chromatogr B Analyt Technol Biomed Life Sci 2008; 870:55-62; PMID:18567545; http://dx.doi.org/10.1016/j.jchromb.2008.05. 045.
70. Wang W, Vlasak J, Li Y, Pristatsky P, Fang Y, Pittman T, et al. Impact of methionine oxidation in human IgG1 Fc on serum half-life of monoclonal antibodies. Mol Immunol 2011; 48:860-6; PMID:21256596; http://dx.doi.org/10. 1016/j.molimm.2010.12.009.