Recombinant porcine HGF (NK1) protein (Qk061)

Recombinant porcine HGF protein is a potent, high-purity NK1 isoform of porcine hepatocyte growth factor (HGF). The porcine recombinant protein HGF differs from human HGF at several key amino acid residues that are predicted to affect bioactivity and receptor binding. Use of species-specific HGF growth factor will facilitate media optimization for cellular agriculture (cultivated meat) and veterinary applications. Also available is Qk060 bovine HGF (NK1). The highly scalable animal origin-free manufacture and enhanced bioactivity make this suitable for chemically-defined media and reproducible scale-up.

Qk061 is a 20 kDa naturally occurring isoform of HGF, animal origin-free and carrier-protein free.

Orders are typically shipped same or next day (except Friday).
Easy world-wide ordering, direct or through our distributors.

1000µg will be despatched as 2 x 500µg

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For any questions, please email orders@qkine.com

Summary

  • High purity recombinant porcine HGF Protein (Uniprot: A0A3Q1LSS6)

  • >98%, by SDS-PAGE quantitative densitometry

  • 20 kDa

  • Expressed in E. coli

  • Animal origin-free (AOF) and carrier protein-free.

  • Manufactured in our Cambridge, UK laboratories

  • Lyophilized from acetonitrile, TFA

  • Resuspend in 10mM HCl at >100 µg/ml (provided with protein and free of charge), prepare single use aliquots, add carrier protein if desired and store frozen at -20°C or -80°C

Featured applications

  • Differentiation of iPSCs to hepatocyte-like cells

  • iPSC and primary cell differentiation to muscle

  • Cellular agriculture and cultivated meat cell culture media optimization

Hepatopoietin-A, Scatter factor, SF

Bioactivity

Bioactivity of porcine hepatocyte growth factor (Qk061)

HGF NK1 activity is determined using the Promega serum response element luciferase reporter assay (*) in HEK293T cells. EC50 = 7.0 ng/ml (338 pM). Cells are treated in triplicate with a serial dilution of HGF NK1 for 3 hours. Firefly luciferase activity is measured and normalized to the control Renilla luciferase activity. Data from Qk061 lot #104417. Please note: protein activity has been determined in a standardized assay using human cells. *Promega pGL4.33[luc2P/SRE/Hygro] #E1340

Purity

Porcine HGF Qk061 protein purity in SDS-PAGE

Recombinant porcine HGF protein (Qk061) migrates as a single band at 18 kDa in non-reducing (NR) conditions and 20 kDa upon reduction (R). Purified recombinant protein (3 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptothanol, R) and non-reduced (NR) conditions and stained with Coomassie Brilliant Blue R250. Data from Qk061 batch #104417.

Further quality assays

  • Mass spectrometry: single species with expected mass

  • Analytical reversed-phase: single sharp peak

  • Endotoxin: <0.005 EU/μg protein (below level of detection)

  • Recovery from stock vial:  >95%

We are a company founded and run by scientists to provide a service and support innovation in stem cell biology and regenerative medicine.  All our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.

Protein background

Hepatocyte growth factor (HGF) regulates cell growth, cell motility, and morphogenesis and plays a central role in angiogenesis, tumorogenesis, and tissue regeneration. HGF binds and activates the receptor tyrosine kinase, c-Met, activating PI3K/AKT, FAK, JNK, and ERK1/2 signaling [1].

HGF is secreted as a single inactive polypeptide and is cleaved by serine proteases into a 62-kDa heavy-chain and 32-36-kDa light-chain. A disulfide bond between the heavy and light chains produces the active, heterodimeric molecule [2]. Alternative splicing of HGF produces multiple transcript variants encoding different isoforms. The NK1 isoform is the smallest naturally occurring splice variant, comprising the N-terminus and first kringle domain [3].

Bovine, porcine and human HGF are all members of the hepatocyte growth factor (HGF) family. While the sequence of porcine HGF is similar to human HGF, there are differences in structure and bioactivity. Porcine HGF is more resistant to protease digestion. Additionally, porcine HGF appears to be more potent than its counterparts when it comes to inducing muscle cell proliferation and differentiation. These findings demonstrate that porcine HGF could have important implications for cellular agriculture applications as it may provide a faster route towards creating tissues with improved regenerative qualities.

Qkine HGF (NK1) recombinant protein is produced in an animal-free microbial system that is compatible with production at scale. We are expanding our range of innovative growth factors for cellular agriculture and cultivated meat media development to support advances in the field, please reach out to cellag@qkine.com to discuss collaboration opportunities.

[1] Organ, Shawna Leslie, and Ming-Sound Tsao. ‘An Overview of the C-MET Signaling Pathway’. Therapeutic Advances in Medical Oncology, vol. 3, no. 1 Suppl, Nov. 2011, pp. S7–19. https://doi.org/10.1177/1758834011422556.

[2] Oliveira, Alexandre G., et al. ‘The Role of Hepatocyte Growth Factor (HGF) in Insulin Resistance and Diabetes’. Frontiers in Endocrinology, vol. 0, 2018. Frontiers, https://doi.org/10.3389/fendo.2018.00503.

[3] Jakubczak, John L., et al. ‘NK1, a Natural Splice Variant of Hepatocyte Growth Factor/Scatter Factor, Is a Partial Agonist In Vivo’. Molecular and Cellular Biology, vol. 18, no. 3, Mar. 1998, pp. 1275–83. doi: 10.1128/mcb.18.3.1275

Species-specificity of HGF is predicted to impact bioactivity

Porcine HGF differs from human and bovine HGF at several key amino acid residues that are predicted to affect bioactivity and receptor binding. Clustal Alignment showing sequence variation between HGF NK1 from different species. Note the lack of structural conservation at key residues, which is predicted to impact receptor binding and activation.

Sequence alignment of porcine, bovine and human HGF NK1 showing amino acid conservation
  • Recombinant human HGF (NK1) protein
  • Qkine HGF (NK1) protein

Our products are for research use only and not for diagnostic or therapeutic use.  Products are not for resale.

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