Skip to main content
SpringerLink
Log in

Analysis of Somatropin by Double-Injection Capillary-Zone Electrophoresis in Polybrene/Chondroitin Sulfate A Double-Coated Capillaries

  • Protocol
  • First Online:
Book cover Capillary Electrophoresis of Proteins and Peptides

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1466))

  • 1371 Accesses

Abstract

Purity determination of somatropin as a recombinant protein is important to ensure its safety and quality. This is carried out by capillary zone electrophoresis in double-injection mode using polybrene/chondroitin sulfate A double-coated capillaries. Modification of the capillary wall eliminates protein-wall interactions which results in improved accuracy and precision of the determinations. In the double-injection mode two somatropin samples are analyzed within a single electrophoretic run. Prior to the second injection, the first injected plug is electrophoresed for a predetermined time period in order to adjust the inter-plug distance. Here, the principle for the separation of somatropin charge variants is described.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Ameeta M, Hindmarsh PC (2002) The use of somatropin (recombinant growth hormone) in children of short stature Pediatric Drugs 4:37–47

    Google Scholar 

  2. Amini A (2013) Separation of somatropin charge variants by multiple-injection CZE with polybrene/chondroitin sulfate A double-coated capillaries. J Sep Sci 36:2686–5690

    Article  CAS  PubMed  Google Scholar 

  3. Doherty EAS, Meagher RJ, Albarghouthi MN, Barron AE (2003) Microchannel wall coatings for protein separations by capillary and chip electrophoresis. Electrophoresis 24:34–54

    Article  PubMed  Google Scholar 

  4. Green JS, Jorgenson JW (1989) Minimizing asorption of proteins on fused silica in capillary zone electrophoresis by the addition of alkali metal salts to the buffers. J Chromatogr A 478:63–70

    CAS  Google Scholar 

  5. Towns JK, Regnier FE (1992) Impact of polycation adsorption on efficiency and electroosmotically driven transport in capillary electrophoresis. Anal Chem 64:2473–2478

    Article  CAS  Google Scholar 

  6. Schwer C, Kenndler E (1991) Ectrophoresis in fused-silica capillaries: the influence of organic solvents on the electroosmotic velocity and the zeta potential. Anal Chem 63:1801–1807

    Google Scholar 

  7. Hjertén S (1985) High-performance electrophoresis: elimination of electroendosmosis and solute adsorption). J Chromatogr 347:191–198

    Article  Google Scholar 

  8. Lauer HH, McManigill D (1986) Capillary zone electrophoresis of proteins in untreated fused silica tubing. Anal Chem 58:166–170

    Article  CAS  Google Scholar 

  9. Chiari M, Damin F, Melis A, Conssoni R (1998) Separation of oligonucleotides and DNA fragments by capillary electrophoresis in dynamically and permanently coated capillaries, using a copolymer of acrylamide and β-d-glucopyranoside as a new low viscosity matrix with high sieving capacity. Electrophoresis 19:3154–3159

    Article  CAS  PubMed  Google Scholar 

  10. Wang Z, Chen Y, Yuan H et al (2000) (Preparation and characterization of calixarene-coated capillaries for capillary electrophoresis. Electrophoresis 21:1620–1624

    Article  CAS  PubMed  Google Scholar 

  11. Dobashi A, Hamada M (1999) Chiral separation of dansyl-DL-amino acids with micellar systems containing copper (II) ion and N-n-dodecyl-L-proline in electrokinetic capillary chromatography. Electrophoresis 20:2761–2765

    Article  CAS  PubMed  Google Scholar 

  12. Erim FB, Cifuentes A, Poppe H, Kraak JC (1995) Performance of a physically adsorbed high-molecular-mass polyethyleneimine layer as coating for the separation of basic proteins and peptides by capillary electrophoresis. J Chromatogr A 708:356–361

    Article  Google Scholar 

  13. Legaz ME, Pedrosa MM (1996) Effect of polyamines on the separation of ovalbumin glycoforms by capillary electrophoresis. J Chromatogr A 719:159–170

    Article  CAS  PubMed  Google Scholar 

  14. Corradini P, Rhomberg A, Corradini C (1994) Electrophoresis of proteins in uncoated capillaries with amines and amino sugars as electrolyte additives. J Chromatogr A 661:305–313

    Article  CAS  Google Scholar 

  15. Olivieri E, Sebastiano R, Citterio A et al (2000) Quantitation of protein binding to the capillary wall in acidic, isoelectric buffers and means for minimizing the phenomenon. J Chromatogr A 894:273–280

    Article  CAS  Google Scholar 

  16. Linder H, Helliger W, Sarg B, Meraner C (1995) (Effect of buffer composition on the migration order and separation of histone H1 subtypes. Electrophoresis 16:604–610

    Article  Google Scholar 

  17. Kleemiss MH, Gilges M, Schomburg G (1993) Capillary electrophoresis of DNA restriction fragments with solutions of entangled polymers. Electrophoresis 14:515–522

    Article  CAS  PubMed  Google Scholar 

  18. Towns JK, Regnier FE (1991) Capillary electrophoretic separations of proteins using nonionic surfactant coatings. Anal Chem 63:1126–1132

    Article  CAS  PubMed  Google Scholar 

  19. Salmanowicz BP (1995) Capillary electrophoresis of seed albumins from vicia species using uncoated and surface-modified fused silica capillaries. Chromatographia 41:99–106

    Article  CAS  Google Scholar 

  20. Oneill K, Shao XW, Zhao ZX et al (1994) Capillary electrophoresis of nucleotides on ucon-coated fused silica columns. Anal Biochem 222:185–189

    Article  CAS  Google Scholar 

  21. Chen SH, Tzeng RT (1999) Polymer solution-filled column for the analysis of antisense phosphorothioates by capillary electrophoresis. Electrophoresis 20:547–554

    Article  CAS  PubMed  Google Scholar 

  22. Iki N, Yeung ES (1996) Non-bonded poly(ethylene oxide) polymer-coated column for protein separation by capillary electrophoresis. J Chromatogr A 731:273–282

    Article  CAS  Google Scholar 

  23. Tsuda T (1987) Modification of electroosmotic flow with cetyltrimethylammonium bromide in capillary zone electrophoresis. J Sep Sci 10:622–624

    CAS  Google Scholar 

  24. Jiang T-F, Gu Y-L, Liang B et al (2003) Dynamically coating the capillary with 1-alkyl-3-methylimidazolium-based ionic liquids for separation of basic proteins by capillary electrophoresis. Anal Chim Acta 479:249–254

    Article  CAS  Google Scholar 

  25. Yao YJ, Li SFY (1991) Capillary zone electrophoresis of basic proteins with chitosan as a capillary modifier. J Chromatogr A 663: 97–104

    Article  Google Scholar 

  26. Cobb KA, Dolnik V, Novotny M (1990) Electrophoretic separations of proteins in capillaries with hydrolytically-stable surface structures. Anal Chem 62:2478–2483

    Article  CAS  PubMed  Google Scholar 

  27. Nashabeh W, El Rassi Z (1991) Capillary zone electrophoresis of proteins with hydrophilic fused-silica capillaries. J Chromatogr A 559:367–383

    Article  CAS  Google Scholar 

  28. Smith JT, El Rassi Z (1993) Capillary zone electrophoresis of biological substances with fused silica capillaries having zero or constant electroosmotic flow. Electrophoresis 14:396–406

    Article  CAS  PubMed  Google Scholar 

  29. McCormick RM (1998) Capillary zone electrophoretic separation of peptides and proteins using low pH buffers in modified silica capillaries. Anal Chem 60:2322–2328

    Article  Google Scholar 

  30. Yao XW, Wu D, Regnier FE (1993) Manipulation of electroosmotic flow in capillary electophoresis. J Chromatogr A 636:21–29

    Article  CAS  Google Scholar 

  31. Shen Y, Smith RD (2000) High-resolution capillary isoelectric focusing of proteins using highly hydrophilic-substituted cellulose-coated capillaries. J Microcol 12:135–141

    Article  CAS  Google Scholar 

  32. Hjertén S, Kubo K (1993) A new type of pH- and detergent-stable coating for elimination of electroendosmosis and adsorption in (capillary) electrophoresis. Electrophoresis 14:390–395

    Article  PubMed  Google Scholar 

  33. Chiari M, Coriotti L, Crini G, Morcellet M (1999) Poly(vinylamine)-coated capillaries with reversed electroosmotic flow for the separation of organic anions. J Chromatogr A 836:81–91

    Article  CAS  Google Scholar 

  34. Belder D, Deege A, Husmann H et al (2001) Cross-linked poly(vinyl alcohol) as permanent hydrophilic column coating for capillary electrophoresis. Electrophoresis 2001(22):3813–3818

    Article  Google Scholar 

  35. Bendahl L, Hansen SH, Gammelgaard B (2001) Capillaries modified by noncovalent anionic polymer adsorption for capillary zone electrophoresis, micellar electrokinetic capillary chromatography and capillary electrophoresis mass spectrometry. Electrophoresis 22:2565–2573

    Article  CAS  PubMed  Google Scholar 

  36. Graul TW, Schlenoff JB (1999) Capillaries modified by polyelectrolyte multilayers for electrophoretic separations. Anal Chem 71:4007–4013

    Article  CAS  Google Scholar 

  37. Aguilar C, Hofte AJP, Tjaden UR, van der Greef J (2001) Analysis of histones by on-line capillary zone electrophoresis–electrospray ionisation mass spectrometry. J Chromatogr A 926:57–67

    Article  CAS  PubMed  Google Scholar 

  38. Busnel JM, Desoroix S, Le Saux T et al (2006) Protein tryptic digests analyzed by carrier ampholyte-based capillary electrophoresis coupled to ESI-MS. Electrophoresis 27:1481–1488

    Article  CAS  PubMed  Google Scholar 

  39. Wahl JH, Gale DC, Smith RD (1994) Sheathless capillary electrophoresis-electrospray ionization mass spectrometry using 10 μm I.D. capillaries: analyses of tryptic digests of cytochrome c. J Chromtogr A 659:217–222

    Article  CAS  Google Scholar 

  40. Katayama H, Ishihama Y, Asakawa N (1998) Stable cationic capillary coating with successive multiple ionic polymer layers for capillary electrophoresis. Anal Chem 70:5272–5277

    Article  CAS  PubMed  Google Scholar 

  41. Amini A, Olofsson IM (2004) Analysis of calcitonin and its analogues by capillary zone electrophoresis and matrix-assisted laserdesorption ionization time-of-flight mass spectrometry. J Sep Sci 27:675–685

    Article  CAS  PubMed  Google Scholar 

  42. Cordova E, Gao J, Whitesides GM (1997) Noncovalent polycationic coatings for capillaries in capillary electrophoresis of proteins. Anal Chem 69:1370–1379

    Article  CAS  PubMed  Google Scholar 

  43. Katayama H, Ishihama Y, Asakawa N (1998) Stable capillary coating with successive multiple ionic polymer layers. Anal Chem 70:5254–5260

    Google Scholar 

  44. Busch MHA, Kraak JC, Poppe H (1995) Cellulose acetate-coated fused-silica capillaries for the separation of proteins by capillary zone electrophoresis. J Chromatogr A 695:287–296

    Article  CAS  Google Scholar 

  45. Haselberg R, Brinks V, Hawe A, de Jong GJ (2011) Capillary electrophoresis-mass spectrometry using non-covalently coated capillaries for the analysis of biopharmaceuticals. Anal Bioanal Chem 400:295–303

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Catai JR, Toraňo JS, Jongen PMJM et al (2007) Analysis of recombinant human growth hormone by capillary electrophoresis with bilayer-coated capillaries using UV and MS detection. J Chromatogr B 852:160–166

    Article  CAS  Google Scholar 

  47. Bateman KP, White RL, Thibault P (1997) Disposable emitters for on-line capillary zone electrophoresis/nanoelectrospray mass spectrometry. Rapid Commun Mass Spectrom 11:307–315

    Article  CAS  Google Scholar 

  48. Bao JJ (2000) Separation of proteins by capillary electrophoresis using epoxy based hydrophilic coating. J Liq Chro Rel Technol 23:61–78

    Article  CAS  Google Scholar 

  49. Melanson EM, Baryla NE, Lucy CA (2000) Double-chained surfactants for semipermanent wall coatings in capillary electrophoresis. Anal Chem 72:4110–4114

    Article  CAS  PubMed  Google Scholar 

  50. Van Tassel PR, Miras D, Hagege A et al (1996) Control of protein adsorption in capillary electrophoresis via an irreversibly bound protein coating. J Colloid Interface Sci 183:269–273

    Article  Google Scholar 

  51. Lucy CA, MacDonal AM, Gulcev MD (2008) Non-covalent capillary coatings for protein separations in capillary electrophoresis. J Chromatogr A 1184:81–105

    Article  CAS  PubMed  Google Scholar 

  52. Amini A (2014) Double injection capillary electrophoresis for the identification of analytes. Electrophoresis 35:2915–2921

    Article  CAS  PubMed  Google Scholar 

  53. Amini A, Lodén H, Pettersson C, Arvidsson T (2008) Principles for different modes of multipleinjection CZE. Electrophoresis 29:3952–3958

    Article  CAS  PubMed  Google Scholar 

  54. Dupin P, Galinou F, Bayol A (1995) Analysis of recombinant human growth hormone and its related impurities by capillary electrophoresis. J Chromatogr A 707:396–400

    Article  CAS  Google Scholar 

  55. Somatropin for injection. European Pharmacopoeia, 8th Edition 2016 (8.8) (01/2008:0952)

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Medical Products Agency (MPA), Box 26, Dag Hammarskjölds väg 42, Uppsala, 751 03, Sweden

    Ahmad Amini

  2. Division of Analytical Pharmaceutical Chemistry, Biomedical Centre, Uppsala University, Box 574, Uppsala, 751 23, Sweden

    Ahmad Amini

Authors
  1. Ahmad Amini
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ahmad Amini .

Editor information

Editors and Affiliations

  1. Institut Galien Paris-Sud, UMR 8612, Protein and Nanotechnology in Analytical Science (PNAS), CNRS, Univ. Paris-Sud, Université Paris-Saclay, Châtenay-Malabry, France

    Nguyet Thuy Tran

  2. Institut Galien Paris-Sud, UMR 8612, Protein and Nanotechnology in Analytical Science (PNAS), CNRS, Univ. Paris-Sud, Université Paris-Saclay, Châtenay-Malabry, France

    Myriam Taverna

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer Science+Business Media New York

About this protocol

Cite this protocol

Amini, A. (2016). Analysis of Somatropin by Double-Injection Capillary-Zone Electrophoresis in Polybrene/Chondroitin Sulfate A Double-Coated Capillaries. In: Tran, N., Taverna, M. (eds) Capillary Electrophoresis of Proteins and Peptides. Methods in Molecular Biology, vol 1466. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-4014-1_8

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-4014-1_8

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-4012-7

  • Online ISBN: 978-1-4939-4014-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation