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Advances on Size Exclusion Chromatography and Applications on the Analysis of Protein Biopharmaceuticals and Protein Aggregates: A Mini Review

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Abstract

Biopharmaceutical drugs are large, complex protein molecules derived from living cells. The characterization of the protein-based products is a complicated and challenging task necessary to guarantee their efficacy and safety. A major concern in manufacturing protein biopharmaceuticals is their propensity to form aggregates. The separation of proteins by size-exclusion chromatography (SEC) is a well-established method for the characterization of macromolecules and their aggregates. The demand for increased resolution and faster analysis in SEC has led to the development of small, more rigid fully porous particles and superficially porous particles with different pore sizes. The coupling of SEC with three or four detectors has enhanced protein characterization by generating accurate molecular weights (absolute determination) and by giving insight into the degradation pathways and products associated with aggregation. The intent of this review is to report the continuous developments of SEC stationary phases and their applications on the characterization of intact proteins, their variants or aggregates and protein complexes. Thus, the most recent applications of SEC with the focus on proteins and protein aggregates are reported and discussed.

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References

  1. Dimitrov DS (2012) In: Voynov V, Caravella JA (eds) Methods mol biol. Springer, New York

    Google Scholar 

  2. Strohl WR, Knight MD (2009) Curr Opin Biotechnol 20:668–672

    Article  CAS  PubMed  Google Scholar 

  3. Mahler HC, Friess W, Grauschopf U, Kiese S (2009) J Pharm Sci 98:2909–2934

    Article  CAS  PubMed  Google Scholar 

  4. Rathore N, Rajan RS (2008) Biotechnol Prog 24:504–514

    Article  CAS  PubMed  Google Scholar 

  5. Manning MC, Chou DK, Murphy BM, Payne RW, Katayama DS (2010) Pharm Res 27:544–575

    Article  CAS  PubMed  Google Scholar 

  6. Patel J, Kothari R, Tunga R, Ritter NM, Tunga BS (2011) BioProcess Int 1:20–31

    Google Scholar 

  7. Staub A, Guillarme D, Schappler J, Veuthey JL, Rudaz S (2011) J Pharm Biomed Anal 55:810–822

    Article  CAS  PubMed  Google Scholar 

  8. Berkowitz SA, Engen JR, Mazzeo JR, Jones GB (2012) Nat Rev Drug Discov 11:527–540

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Kaltashov IA, Bobst CE, Abzalimod RR, Wang G, Baykal B (2012) Wang S Biotechnol Adv 30:210–222

    Article  CAS  PubMed  Google Scholar 

  10. Oliva A, Fariña JB, Llabrés M (2007) Curr Pharm Anal 3:230–248

    Article  CAS  Google Scholar 

  11. Parr MK, Montacir O, Montacir H (2016) J Pharm Biomed Anal 130:366–389

    Article  CAS  PubMed  Google Scholar 

  12. Zhang H, Cui W, Gross ML (2014) FEBS Lett 588:308–317

    Article  CAS  PubMed  Google Scholar 

  13. Zhang Z, Pan H, Chen X (2009) Mass Spectrom Rev 28:147–176

    Article  CAS  PubMed  Google Scholar 

  14. Fekete S, Veuthey JL, Guillarme D (2015) LCGC Special issues Oct 02: 8–15

  15. Bobály B, Sipkó E, Fekete J (2016) J Chromatogr B 1032:3-2

    Article  CAS  Google Scholar 

  16. Fekete S, Veuthey JL, Guillarme D (2017) J Pharm Biomed Anal 141:59–69

    Article  CAS  PubMed  Google Scholar 

  17. Sandra K, Vandenheede I, Sandra P (2014) J Chromatogr A 1335:81–103

    Article  CAS  PubMed  Google Scholar 

  18. Oliva A, Llabres M, Farina JB (2001) J Pharm Biomed Anal 25:833–841

    Article  CAS  PubMed  Google Scholar 

  19. Hong P, Koza S, Bouvier ESP (2012) J Liq Chromatogr RT 35:2923–2950

    CAS  Google Scholar 

  20. Patten PA, Schellekens H (2003) In: Brown F, Mire-Sluis AR (eds) The immunogenicity of biopharmaceuticals: lessons learned and consequences for protein drug development. Karger, Basel

    Google Scholar 

  21. Rosenberg AS (2006) AAPS J 8:E501–E507

    Article  PubMed  PubMed Central  Google Scholar 

  22. Philo JS (2009) Curr Pharm Biotechnol 10:359–372

    Article  CAS  PubMed  Google Scholar 

  23. Narhi LO, Schmit J, Bechtold-Peters K, Sharma D (2012) J Pharm Sci 101:493–498

    Article  CAS  PubMed  Google Scholar 

  24. Hamrang Z, Rattray NJW, Pluen A (2013) Trends Biotechnol 31:448–458

    Article  CAS  PubMed  Google Scholar 

  25. Smulders R, Koll H, Smith B, Bassarab S, Seidl A, Hainzl O, Jiskoot W (2011) Pharm Res 28:920–933

    Article  CAS  PubMed  Google Scholar 

  26. Khodabandehloo A, Chen DD (2017) Bioanalysis 9:313–326

    Article  CAS  PubMed  Google Scholar 

  27. Gandhi AV, Pothecary MR, Bain DL, Carpenter JF (2017) J Pharm Sci 106:2178–2186

    Article  CAS  PubMed  Google Scholar 

  28. Fekete S, Beck A, Veuthey JL, Guillarme D (2014) J Pharm Biomed Anal 101:161–173

    Article  CAS  PubMed  Google Scholar 

  29. Uliyanchenko E (2014) Anal Bioanal Chem 406:6087–6094

    Article  CAS  PubMed  Google Scholar 

  30. Striegel A, Yau WW, Kirkland JJ, Bly DD (2009) Modern size-exclusion liquid chromatography: practice of gel permeation and gel filtration chromatography, 2nd edn. Wiley, New York

    Book  Google Scholar 

  31. Wu CS (2003) Handbook of size-exclusion chromatography and related techniques. Marcel Dekker, New York

    Book  Google Scholar 

  32. Saunders GD, Barth HG (2012) LCGC North Am 30:544–563

    Google Scholar 

  33. Hudalla CJ, Alden B, Walter T, Walsh D, Bouvier E, Iraneta PC, Lawrence N, Wyndham K (2012) LCGC North Am 30:20–29

    Google Scholar 

  34. Wyndham KD, Walter TH, Iraneta PC, Alden BA, Bouvier ESP, Hudalla CJ, Lawrence NL, Walsh D (2012) LCGC North Am 30(S4):20–29

    Google Scholar 

  35. Unger KK, Liapis AI (2012) J Sep Sci 35:1201–1212

    Article  CAS  PubMed  Google Scholar 

  36. Wagner BM, Schuster SA, Boyes BE, Kirkland JJ (2012) J Chromatogr A 1264:22–30

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Fekete S, Ganzler K, Guillarme D (2013) J Pharm Biomed Anal 78–79:141–149

    Article  CAS  PubMed  Google Scholar 

  38. Schure MR, Moran RE (2017) J Chromatogr A 1480:11–19

    Article  CAS  PubMed  Google Scholar 

  39. Pirok BWJ, Breuer P, Serafine JM, Hoppe SJM, Chitty M, Welch E, Farkas T, van der Wal S, Peters R, Schoenmakers PJ (2017) J Chromatogr A 1486:96–102

    Article  CAS  PubMed  Google Scholar 

  40. Wagner BM, Schuster SA, Boyes BE, Shieldsa TJ, Miles WL, Haynes MJ, Moran RE, Kirkland JJ, Schure MR (2017) J Chromatogr A 1489:75–85

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Fekete S, Oláh E, Fekete J (2012) J Chromatogr A 1228:57–71

    Article  CAS  PubMed  Google Scholar 

  42. Bouvier ESP, Koza SM (2014) Trends Anal Chem 63:85–94

    Article  CAS  Google Scholar 

  43. Fekete S, Guillarme D (2014) Trends Anal Chem 63:76–84

    Article  CAS  Google Scholar 

  44. Yang R, Tang Y, Zhang B, Lu X, Liu A, Zhang YZ (2015) J Pharm Biomed Anal 109:52–61

    Article  CAS  PubMed  Google Scholar 

  45. Goyon A, Beck A, Colas O, Sandra K, Guillarme D, Fekete S (2017) J Chromatogr A 1498:80–89

    Article  CAS  PubMed  Google Scholar 

  46. Goyon A, Guillarme D, Fekete S (2017) J Pharm Biomed Anal 135:50–60

    Article  CAS  PubMed  Google Scholar 

  47. Krull IS, Rathore AS (2015) LCGC Europe 28:54–58

    Google Scholar 

  48. Haberger Taraban MB, DePaz RA, Lobo B, Yu YB (2017) Anal Chem 89:5494–5502

    Article  CAS  Google Scholar 

  49. Leiss M, Heidenreich AK, Pester O, Hafenmair G, Hook M, Bonnington L, Wegele H, Haindl M, Reusch R, Bulau P (2016) Mabs 8:331–339

    Article  CAS  PubMed  Google Scholar 

  50. Striegel AM (2005) Anal Chem 77:104A–113A

    Article  CAS  Google Scholar 

  51. Fekete S, Guillarme D, Sandra S, Sandra K (2016) Anal Chem 88:480–507

    Article  CAS  PubMed  Google Scholar 

  52. Liu J, Chen X, Fan L, Deng X, Fai Poon H, Tan WS, Liu X (2015) Biotechnol Lett 37:1371–1377

    Article  CAS  PubMed  Google Scholar 

  53. Liu F, Reinmuth-Selzle K, Lai S, Weller MG, Pöschl U, Kampf CJ (2017) J Chromatogr A 1495:76–82

    Article  CAS  PubMed  Google Scholar 

  54. Tazi LM, Shiranthi J (2016) J Chromatogr B 1011:89–93

    Article  CAS  Google Scholar 

  55. Wang H, Levi MS, Del Grosso AV, McCormick WM, Bhattacharyya LJPBA (2017) J Pharm Biomed Anal 138:330–343

    Article  CAS  PubMed  Google Scholar 

  56. De Vos J, Kaal ER, Swart R, Baca M, Vander Heyden Y, Eeltink S (2016) J Sep Sci 39:689–695

    Article  CAS  PubMed  Google Scholar 

  57. Goyon A, D’Atria V, Bobaly B, Wagner-Rousset E, Beck A, Fekete S, Guillarme D (2017) J Chromatogr B 1058:73–84

    Article  CAS  Google Scholar 

  58. Jin Y, Wei L, Cai W, Lin Z, Wu Z, Peng Y, Kohmoto T, Moss RL, Ge Y (2017) Anal Chem 89:4922–4930

  59. Cai W, Tucholski T, Chen B, Alpert AJ, McIlwain S, Kohmoto T, Jin S, Ge Y (2017) Anal Chem 89:5467–5475

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Makarov AA, Helmyet R (2016) J Chromatogr A 1431:224–230

    Article  CAS  PubMed  Google Scholar 

  61. Pierson NA, Makarov AA, Strulson CA, Mao Y, Mao B (2017) J Chromatogr A 1496:51–57

    Article  CAS  PubMed  Google Scholar 

  62. Vemula S, Vemula S, Dedaniya A, Ronda SR (2016) Anal Bioanal Chem 408:217–229

    Article  CAS  PubMed  Google Scholar 

  63. Li W, Yang B, Zhou D, Xu J, Li W, Suen W-C (2017) J Chromatogr B 1048:121–129

    Article  CAS  Google Scholar 

  64. Beck A, Wagner-Rousset E, Ayoub D, Van Dorsselaer A, Sanglier-Cianferani S (2013) Anal Chem 85:715–736

    Article  CAS  PubMed  Google Scholar 

  65. Gervais D, Downer A, King D, Kanda P, Foote N, Smith S (2017) J Pharm Biomed Anal 139:215–220

    Article  CAS  PubMed  Google Scholar 

  66. Espinosa-de la Garza CE, Miranda-Hernandez MP, Acosta-Flores L, Perez NO, Flores-Ortiz LF, Medina-Rivero E (2015) J Sep Sci 38:1537–1543

    Article  CAS  PubMed  Google Scholar 

  67. Miranda-Hernandez MP, Elba R, Valle-González ER, Ferreira-Gómez D, Pérez NO, Flores-Ortiz LF, Medina-Rivero E (2016) Anal Bioanal Chem 408:1523–1530

    Article  CAS  PubMed  Google Scholar 

  68. Svedlund FL, Eda I, Altiok EI, Healy KE (2016) Biomacromolecules 17:3162–3171

    Article  CAS  PubMed  Google Scholar 

  69. Singla A, Bansal R, Joshi V, Rathore AS (2016) AAPS J 18:689–702

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  70. Bickel F, Herold EM, Signes A, Romeijn S, Jiskoot W, Kiefer H (2016) Eur J Pharm Biopharm 107:310–320

    Article  CAS  PubMed  Google Scholar 

  71. Bourrelle-Langlois M, Morrow G, Finet S, Tanguay RM (2016) PLoS One 11:1–21

    Article  CAS  Google Scholar 

  72. Yang Y, Li H, Zhengjun LZ, Zhanga Y, Zhanga S, Chena Y, Yua M, Ma G, Su Z (2015) Vaccine 33:1143–1150

    Article  CAS  PubMed  Google Scholar 

  73. Vajda J, Weber D, Brekel D, Hundt B, Müller E (2016) J Chromatogr A 1465:117–125

    Article  CAS  PubMed  Google Scholar 

  74. Ladd-Effio C, Stefan A, Oelmeier SA, Hubbuch J (2016) Vaccine 34:1259–1267

    Article  CAS  PubMed  Google Scholar 

  75. Lancaster C, Rustandi RR, Pannizzo P, Ha S (2016) Methods Mol Biol 1476:279–287

    Article  PubMed  Google Scholar 

  76. Gargani-Weisbjerg PL, Bjerg Caspersen M, Cook K, Van De Weert M (2015) J Pharm Sci 104:548–556

    Article  CAS  Google Scholar 

  77. Malaby AW, Srinivas Chakravarthy S, Thomas C, Irving TC, Sagar V, Kathuria SV, Bilsel O, Lambright DG (2015) J Appl Crystallogr 48:1102–1113

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  78. Jordan A, Jacques M, Merrick C, Devos J, Forsyth VT, Porcar L, Martel A (2016) J Appl Crystallogr 49:2015–2020

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  79. Gjoka X, Schofield M, Cvetkovic A, Gantier R (2014) J Chromatogr B 972:48–52

    Article  CAS  Google Scholar 

  80. Yearley EJ, Godfrin PD, Perevozchikova T, Zhang H, Falus P, Porcar L, Nagao M, Curtis JE, Gawande P, Taing R, Isidro E, Zarraga IE, Wagner NJ, Yun Liu Y (2014) Biophys J 106:1763–1770

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  81. Ojala F, Sellberg A, Hansen TH, Broberg Hansen E, Nilsson B (2015) Biotechnol J 10:1814–1821

    Article  CAS  PubMed  Google Scholar 

  82. Hernandez-Jimenez J, Salmeron-García A, Cabeza J, Velez C, Capitan-Vallvey LF, Navas N (2016) J Pharm Sci 105:1405–1418

    Article  CAS  PubMed  Google Scholar 

  83. Kukrer B, Vasco Filipe V, van Duijn E, Kasper PT, Vreeken RJ, Heck AJR, Jiskoot W (2010) Pharm Res 27:2197–2204

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  84. Wang G, Johnson AJ, Kaltashov IA (2012) Anal Chem 84:1718–1724

    Article  CAS  PubMed  Google Scholar 

  85. Muneeruddin K, Thomas JJ, Salinas PA, Kaltashov IA (2014) Anal Chem 86:10692–10699

    Article  CAS  PubMed  Google Scholar 

  86. Li Y, Gu C, Gruenhagen J, Zhang K, Yehl P, Chetwyn NP, Medley CD (2015) J Chromatogr A 1393:81–88

    Article  CAS  PubMed  Google Scholar 

  87. Hildebrandt C, Mathaes R, Saedler R, Winter G (2016) J Pharm Sci 105:1059–1065

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Drug Sciences, University of Pavia, Viale Taramelli 12, 27100, Pavia, Italy

    Gloria Brusotti, Enrica Calleri, Raffaella Colombo, Gabriella Massolini, Francesca Rinaldi & Caterina Temporini

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  1. Gloria Brusotti
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  2. Enrica Calleri
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  3. Raffaella Colombo
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  4. Gabriella Massolini
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  5. Francesca Rinaldi
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  6. Caterina Temporini
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Correspondence to Gabriella Massolini.

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Published in the topical collection Peptide and Protein Analysis with Debby Mangelings and Gerhard K. E. Scriba as editors.

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Brusotti, G., Calleri, E., Colombo, R. et al. Advances on Size Exclusion Chromatography and Applications on the Analysis of Protein Biopharmaceuticals and Protein Aggregates: A Mini Review. Chromatographia 81, 3–23 (2018). https://doi.org/10.1007/s10337-017-3380-5

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  • DOI: https://doi.org/10.1007/s10337-017-3380-5

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