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Production of recombinant human type I procollagen homotrimer in the mammary gland of transgenic mice

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Abstract

The large scale production of recombinant collagen for use in biomaterials requires an efficient expression system capable of processing a large (>400 Kd) multisubunit protein requiring post-translational modifications. To investigate whether the mammary gland of transgenic animals fulfills these requirements, transgenic mice were generated containing the αS1-casein mammary gland-specific promoter operatively linked to 37 Kb of the human α1(I) procollagen structural gene and 3′ flanking region. The frequency of transgenic lines established was 12%. High levels of soluble triple helical homotrimeric [(α1)3] type I procollagen were detected (up to 8 mg/ml) exclusively in the milk of six out of 9 lines of lactating transgenic mice. The transgene-derived human procollagen chains underwent efficient assembly into a triple helical structure. Although proline or lysine hydroxylation has never been described for any milk protein, procollagen was detected with these post-translational modifications. The procollagen was stable in mil; minimal degradation was observed. These results show that the mammary gland is capable of expressing a large procollagen gene construct, efficiently assembling the individual polypeptide chains into a stable triple helix, and secreting the intact molecule into the milk.

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  1. Cohesion Technologies, 2500 Faber Place, PaloAlto, CA, 94303, USA

    P. David Toman, Naomi Sakai, Gregory A. Daniels & Richard A. Berg

  2. Pharming BV, Niels Bohrweg, 11‐13, 2333 CALeiden, The Netherlands

    Frank Pieper, Costas Karatzas, Erika Platenburg, Ineke de Wit, Caroline Samuel, Anita Dekker & Gerard J. Platenburg

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  1. P. David Toman
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Toman, P.D., Pieper, F., Sakai, N. et al. Production of recombinant human type I procollagen homotrimer in the mammary gland of transgenic mice. Transgenic Res 8, 415–427 (1999). https://doi.org/10.1023/A:1008959924856

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