Skip to main content
SpringerLink
Log in

Production of recombinant albumin by a herd of cloned transgenic cattle

  • Original Paper
  • Published:
Transgenic Research Aims and scope Submit manuscript

Abstract

Purified plasma derived human albumin has been available as a therapeutic product since World War II. However, cost effective recombinant production of albumin has been challenging due to the amount needed and the complex folding pattern of the protein. In an effort to provide an abundant source of recombinant albumin, a herd of transgenic cows expressing high levels of rhA in their milk was generated. Expression cassettes efficiently targeting the secretion of human albumin to the lactating mammary gland were obtained and tested in transgenic mice. A high expressing transgene was transfected in primary bovine cell lines to produce karyoplasts for use in a somatic cell nuclear transfer program. Founder transgenic cows were produced from four independent cell lines. Expression levels varying from 1–2 g/l to more than 40 g/l of correctly folded albumin were observed. The animals expressing the highest levels of rhA exhibited shortened lactation whereas cows yielding 1–2 g/l had normal milk production. This herd of transgenic cattle is an easily scalable and well characterized source of rhA for biomedical uses.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Alexander MR, Alexander B, Mustion AL, Spector R, Wright CB (1982) Therapeutic use of albumin. JAMA 247:831–833. doi:10.1001/jama.247.6.831

    Article  PubMed  CAS  Google Scholar 

  • Baguisi A, Behboodi E, Melican DT, Pollock JS, Detrempes MM, Cammuso C, Williams JL, Nims SD, Porter CA, Midura P, Palacios MJ, Ayers SL, Denniston RS, Hayes ML, Ziomek CA, Meade HM, Godke RA, Gavin WG, Overström EW, Echelard Y (1999) Production of goats by somatic cell nuclear transfer. Nat Biotechnol 17:456–461. doi:10.1038/8632

    Article  PubMed  CAS  Google Scholar 

  • Behboodi E, Groen W, Destrempes MM, Williams JL, Ohlrichs C, Gavin WG, Broek DM, Ziomek CA, Faber DC, Meade HM, Echelard Y (2001) Transgenic production from in vivo-derived embryos: effect on calf birth weight and sex ratio. Mol Reprod Dev 60:27–37. doi:10.1002/mrd.1058

    Article  PubMed  CAS  Google Scholar 

  • Brophy B, Smolenski G, Wheeler T, Wells D, L’Huillier P, Laible G (2003) Cloned transgenic cattle produce milk with higher levels of beta-casein and kappa-casein. Nat Biotechnol 21:157–162. doi:10.1038/nbt783

    Article  PubMed  CAS  Google Scholar 

  • Chen RF (1967) Removal of fatty acids from serum albumin by charcoal treament. J Biol Chem 242:173–181

    PubMed  CAS  Google Scholar 

  • Chuang VT, Kragh-Hansen U, Otagiri M (2002) Pharmaceutical strategies utilizing recombinant human serum albumin. Pharm Res 19:569–577. doi:10.1023/A:1015396825274

    Article  PubMed  Google Scholar 

  • Chung JH, Whiteley M, Felsenfeld G (1993) A 5′ element of the chicken beta-globin domain serves as an insulator in human erythroid cells and protects. Cell 74:505–514. doi:10.1016/0092-8674(93)80052-G

    Article  PubMed  CAS  Google Scholar 

  • Church GM, Gilbert W (1984) Genomic sequencing. Proc Natl Acad Sci USA 81:1991–1995. doi:10.1073/pnas.81.7.1991

    Article  PubMed  CAS  Google Scholar 

  • Cibelli JB, Stice SL, Golueke PJ, Kane JJ, Jerry J, Blackwell C, Ponce de Leon A, Robl JM (1998) Cloned transgenic calves produced from nonquiescent fetal fibroblasts. Science 280:1256–1258. doi:10.1126/science.280.5367.1256

    Article  PubMed  CAS  Google Scholar 

  • Clark AJ (1998) The mammary gland as a bioreactor: Expression, processing, and production of recombinant proteins. J Mammary Gland Biol 3:337–350. doi:10.1023/A:1018723712996

    Article  CAS  Google Scholar 

  • Cox H, Mead D, Sudbery P, Eland RM, Mannazzu I, Evans L (2000) Constitutive expression of recombinant proteins in the methylotrophic yeast Hansenula polymorpha using the PMA1 promoter. Yeast 30:1191–1203. doi:10.1002/1097-0061(20000930)16:13<1191::AID-YEA589>3.0.CO;2-2

    Article  Google Scholar 

  • Damiani P, Fissore RA, Cibelli JB, Long CR, Balise JJ, Robl JM, Duby RT (1996) Evaluation of developmental competence, nuclear and ooplasmic maturation of calf oocytes. Mol Reprod Dev 45:521–534. doi:10.1002/(SICI)1098-2795(199612)45:4<521::AID-MRD15>3.0.CO;2-Z

    Article  PubMed  CAS  Google Scholar 

  • DiTullio P, Cheng SH, Marshall J, Gregory RJ, Ebert KM, Meade HM, Smith AE (1992) Production of cystic fibrosis transmembrane conductance regulator in the milk of transgenic mice. Biotechnology (NY) 10:74–77. doi:10.1038/nbt0192-74

    Article  CAS  Google Scholar 

  • Drouin R, Harvey P, Richer CL, Rémy-Prince S, Montplaisir S (1987) Human serum albumin as a substitute for foetal calf serum in blast transformation assays and cytogenetic analyses. Cytobios 53:193–207

    Google Scholar 

  • Ebert KM, DiTullio P, Barry CA, Schindler JE, Ayres SL, Smith TE, Pellerin LJ, Meade HM, Denman J, Roberts B (1994) Induction of human tissue plasminogen activator in the mammary gland of transgenic goats. Biotechnology (NY) 12:699–702. doi:10.1038/nbt0794-699

    Article  CAS  Google Scholar 

  • Echelard Y, Destrempes MM, Koster JA, Blackwell C, Groen W, Pollock D, Williams JL, Behboodi E, Pommer J, Meade HM (2002) Production of recombinant human serum albumin in the milk of transgenic cows. Theriogenology 57:779

    Google Scholar 

  • Echelard Y, Ziomek CA, Meade HM (2006) Production of recombinant therapeutic proteins in the milk of transgenic animals. BioPharm Int 19:36–46

    CAS  Google Scholar 

  • Erstad BL, Gales BJ, Rappaport WD (1991) The use of albumin in clinical practice. Arch Intern Med 151:901–911. doi:10.1001/archinte.151.5.901

    Article  PubMed  CAS  Google Scholar 

  • Farran I, Sanchez-Serrano JJ, Medina JF, Prieto J, Mingo-Castel AM (2002) Targeted expression of human serum albumin to potato tubers. Transgenic Res 11:337–346. doi:10.1023/A:1016356510770

    Article  PubMed  CAS  Google Scholar 

  • Finlayson JS (1980) Albumin products. Semin Thromb Hemost 6:85–120

    Article  CAS  Google Scholar 

  • Fleer R, Yeh P, Amelllal N, Maury I, Fournier A, Bacchetta F, Baduel P, Jung G, L’Hote H, Becquart J, Fukuhara H, Mayaux JF (1991) Stable multicopy vectors for high-level secretion of recombinant human serum albumin by Kluyveromyces yeasts. Biotechnology (NY) 9:968–975. doi:10.1038/nbt1091-968

    Article  CAS  Google Scholar 

  • Gutierrez A, Meade H, Ditullio P, Pollock D, Harvey M, Jimenez-Flores R, Anderson G, Murray J, Medrano J (1996) Expression of a bovine kappa-CN cDNA in the mammary gland of transgenic mice utilizing a genomic milk protein gene as an expression cassette. Transgenic Res 5:271–279. doi:10.1007/BF01972881

    Article  PubMed  CAS  Google Scholar 

  • Haselbeck A (2003) Epoetins: differences and their relevance to immunogenicity. Curr Med Res Opin 19:430–432

    Article  PubMed  CAS  Google Scholar 

  • Hennessen W (1980) The clinical uses of albumin: report of an IABS study. Dev Biol Stand 48:49–52

    PubMed  CAS  Google Scholar 

  • Hewick RM, Hunkapiller MW, Hood LE, Dreyer WJ (1981) A gas-liquid solid phase peptide and protein sequenator. J Biol Chem 256:7990–7997

    PubMed  CAS  Google Scholar 

  • Hogan F, Constantini F, Lacy E (1986) Manipulating the mouse embryo: a laboratory manual. Cold Spring Harbor Lab. Press, Plainview

    Google Scholar 

  • Holst N, Bertheussen K, Forsdahl F, Hakonsen MB, Hansen LJ, Nielsen HI (1990) Optimization and simplification of culture conditions in human in vitro fertilization (IVF) and preembryo replacement by serum-free media. J In Vitro Fertil Em 7:47–53. doi:10.1007/BF01133884

    Article  CAS  Google Scholar 

  • Houdebine LM (1994) The production of pharmaceutical proteins from the milk of transgenic animals. J Biotechnol 34:269–287. doi:10.1016/0168-1656(94)90062-0

    Article  PubMed  CAS  Google Scholar 

  • Kobayashi K, Nakamura N, Sumi A, Ohmura T, Yokoyama K (1998) The development of recombinant human serum albumin. Ther Apher 2:257–262. doi:10.1111/j.1744-9987.1998.tb00118.x

    Article  PubMed  CAS  Google Scholar 

  • Laird PW, Zijderveld A, Linders K, Rudnicki MA, Jaenisch R, Berns A (1991) Simplified mammalian DNA isolation procedure. Nucleic Acids Res 19:4293. doi:10.1093/nar/19.15.4293

    Article  PubMed  CAS  Google Scholar 

  • Lanza RP, Cibelli JB, Blackwell C, Cristofalo VJ, Francis MK, Baerlocker GM, Mak J, Schertzer M, Chavez EA, Sawyer N, Lansdorp PM, West MD (2000) Extension of cell life-span and telomere length in animals cloned from senescent somatic cells. Science 288:665–669. doi:10.1126/science.288.5466.665

    Article  PubMed  CAS  Google Scholar 

  • Lawn RM, Adelman J, Bock SC, Franke AE, Houck CM, Najarian RC, Seeburg PH, Wion KL (1981) The sequence of human serum albumin cDNA and its expression in E. coli. Nucleic Acids Res 25:6103–6114. doi:10.1093/nar/9.22.6103

    Article  Google Scholar 

  • Ledford H (2006) The farmyard drug store. Nature 443:16–17. doi:10.1038/443733a

    Article  CAS  Google Scholar 

  • Maga EA, Murray JD (1995) Mammary gland expression of transgenes and the potential for altering the properties of milk. Biotechnology (NY) 13:1452–1457. doi:10.1038/nbt1295-1452

    Article  CAS  Google Scholar 

  • Marth E, Kleinhappl B (2001) Albumin is a necessary stabilizer of TBE-vaccine to avoid fever in children after vaccination. Vaccine 20:532–537. doi:10.1016/S0264-410X(01)00329-2

    Article  PubMed  CAS  Google Scholar 

  • Morimoto Y, Fujimoto S (1985) Albumin microspheres as drug carriers. Crit Rev Ther Drug 2:19–63

    CAS  Google Scholar 

  • Niemann H, Kues WA (2007) Transgenic farm animals: an update. Reprod Fertil Dev 19:762–770. doi:10.1071/RD07040

    Article  PubMed  Google Scholar 

  • Ohtani W, Nawa Y, Takeshima K, Kamuro H, Kobayashi K, Ohmura T (1998) Physicochemical and immunochemical properties of recombinant human serum albumin from Pichia pastoris. Anal Biochem 256:56–62. doi:10.1006/abio.1997.2480

    Article  PubMed  CAS  Google Scholar 

  • Peters T Jr (ed) (1996) All about albumin: biochemistry genetics and medical applications. Academic Press, San Diego

    Google Scholar 

  • Pollock DP, Kutzko JP, Birck-Wilson E, Williams JL, Echelard Y, Meade HM (1999) Transgenic milk as a method for the production of recombinant antibodies. J Immunol Methods 231:147–157. doi:10.1016/S0022-1759(99)00151-9

    Article  PubMed  CAS  Google Scholar 

  • Quirk AV, Geisow MJ, Woodrow JR, Burton SJ, Wood PC, Sutton AD, Johnson RA, Dodsworth N (1989) Production of recombinant human serum albumin from Saccharomyces cerevisiae. Biotechnol Appl Biochem 11:273–287

    PubMed  CAS  Google Scholar 

  • Roberts B, DiTullio P, Vitale J, Hehir K, Gordon K (1992) Cloning of the goat b-casein-encoding gene and expression in transgenic mice. Gene 121:225–262

    Google Scholar 

  • Robertson I, Nelson RE (1998) Certification and identification of the embryo. In: Stringfellow DA, Seidel SM (eds) Manual of the international embryo transfer society, vol 3. International Embryo Transfer Society, Savoy

    Google Scholar 

  • Rudolph NS (1995) Regulatory issues relating to protein production in transgenic animal milk. Genet Eng News 15:16–18

    Google Scholar 

  • Saunders CW, Schmidt BJ, Mallonee RL, Guyer MS (1987) Secretion of human serum albumin from Bacillus subtilis. J Bacteriol 169:2917–2925

    PubMed  CAS  Google Scholar 

  • Shaklai N, Garlick RL, Bunn HF (1984) Nonenzymatic glycosylation of human serum albuman alters its confirmation and function. J Biol Chem 259:3812–3817

    PubMed  CAS  Google Scholar 

  • Shani M, Barash I, Nathan M, Ricca G, Searfoss GH, Dekel I, Faerman A, Givol D, Hurwitz DR (1992) Expression of human serum albumin in the milk of transgenic mice. Transgenic Res 1:195–208. doi:10.1007/BF02524750

    Article  PubMed  CAS  Google Scholar 

  • Sijmons PC, Dekker BM, Schrammeijer B, Verwoerd TC, van den Elzen PJ, Hoekema A (1990) Production of correctly processed human serum albumin in transgenic plants. Biotechnology (NY) 8:217–221. doi:10.1038/nbt0390-217

    Article  CAS  Google Scholar 

  • Soriano P, Montgomery C, Geske R, Bradley A (1991) Targeted disruption of the c-src proto-oncogene leads to osteopetrosis in mice. Cell 64:693–702. doi:10.1016/0092-8674(91)90499-O

    Article  PubMed  CAS  Google Scholar 

  • Stadalnik RC, Kudo M, Eckelman WC, Vera DR (1993) In vivo functional imaging using receptor-binding radiopharmaceuticals; technetium 99 m-galactosyl-neoglycoalbumin as a model. Invest Radiol 28:64–70. doi:10.1097/00004424-199301000-00018

    Article  PubMed  CAS  Google Scholar 

  • Tarelli E, Mire-Sluis A, Tivnann HA, Bolgiano B, Crane DT, Gee C, Lemercinier X, Athayde ML, Sutcliffe N, Corran PH, Rafferty B (1998) Recombinant human albumin as a stabilizer for biological materials and for the preparation of international reference reagents. Biologicals 26:331–346. doi:10.1006/biol.1998.0163

    Article  PubMed  CAS  Google Scholar 

  • Tullis JL (1977) Albumin 1 background and use. JAMA 237:355–360. doi:10.1001/jama.237.4.355

    Article  PubMed  CAS  Google Scholar 

  • van Berkel PHC, Welling MM, Geerts M, van Veen HA, Ravensbergen B, Salaheddine M, Pauwels EKJ, Pierper F, Nuijens JH, Nibbering PH (2002) Large scale production of recombinant human lactoferrin in the milk of transgenic cows. Nat Biotechnol 20:484–487. doi:10.1038/nbt0502-484

    Article  PubMed  CAS  Google Scholar 

  • Wilkes MM, Navickis RJ (2001) Patient survival after human albumin administration a meta-analysis of randomized, controlled trials. Ann Intern Med 135:149–164

    PubMed  CAS  Google Scholar 

  • World Health Organization (1996) Bovine spongiform encephalopathy. Wkly Epidemiol Rec 71:113–116

    Google Scholar 

  • World Health Organization guidelines on transmissible spongiform encephalopathies in relation to biological and pharmaceutical products (2003) World Health Organization, Blood safety and clinical technology department, Health technology and pharmaceuticals cluster Geneva, Switzerland

  • Zawada WM, Cibelli JB, Choi PK, Clarkson ED, Golueke PJ, Witta SE, Bell KP, Kane J, Ponce de Leon FA, Jerry DJ, Robl JM, Freed CR, Stice SL (1998) Somatic cell cloned transgenic bovine neurons for transplantation in parkinsonian rats. Nat Med 4:569–574. doi:10.1038/nm0598-569

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank our colleagues Cathy Blackwell, Dr. Carol Ziomek, Gregory Liposky, Scott Fulton, Christopher Hendry, Debra Trainor, Daniel Couto, France Dumas, Dr. Eszter Birck-Wilson, Dr. Steven Stice, Dr. James Robl, Dr. Volker Lang, and Dr. Wolfram Eichner for their expert advice, support and encouragement at numerous stages of this project. This work was supported in part by Grant 1R44HL078300-01 from the NIH National Heart, Lung and Blood Institute.

Author information

Authors and Affiliations

  1. GTC Biotherapeutics, Inc., 5 Mountain Road, Framingham, MA, 01701-9322, USA

    Yann Echelard, Jennifer L. Williams, Margaret M. Destrempes, Susan A. Overton, Daniel P. Pollock, Karen T. Rapiejko, Esmail Behboodi, Nicholas C. Masiello, William G. Gavin & Harry M. Meade

  2. Trans Ova Genetics, 2938 380th Street, Sioux Center, IA, 51250, USA

    Julie A. Koster, Jerry Pommer & David C. Faber

  3. Genzyme Corp., Cell and Protein Therapeutics, Framingham, MA, 01701-9322, USA

    Scott M. Van Patten

  4. Departments of Animal Science and Physiology, Michigan State University, East Lansing, MI, USA

    Jose B. Cibelli

  5. PACTMER, Andalucia, Spain

    Jose B. Cibelli

Authors
  1. Yann Echelard
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jennifer L. Williams
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Margaret M. Destrempes
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Julie A. Koster
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Susan A. Overton
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Daniel P. Pollock
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Karen T. Rapiejko
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Esmail Behboodi
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Nicholas C. Masiello
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. William G. Gavin
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Jerry Pommer
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Scott M. Van Patten
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. David C. Faber
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Jose B. Cibelli
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Harry M. Meade
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yann Echelard.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Echelard, Y., Williams, J.L., Destrempes, M.M. et al. Production of recombinant albumin by a herd of cloned transgenic cattle. Transgenic Res 18, 361–376 (2009). https://doi.org/10.1007/s11248-008-9229-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11248-008-9229-9

Keywords

Search

Navigation