Journal: Marine Biotechnology (New York, N.y.)

Article Title: Production of Biopharmaceutical Proteins from Scallop Hemocytes as a Bioreactor Using Improved Transfection Conditions

doi: 10.1007/s10126-026-10583-9

Figure Lengend Snippet: Multi-level confirmation of hGH expression in scallop hemocytes. ( A ) Schematic diagrams of the EGFP vector and two hGH vectors tagged with EGFP or V5/His. ( B ) Fluorescence imaging of hGH::EGFP expression in cultured hemocytes (middle). Green and blue signals represent EGFP fluorescence and DAPI-stained nuclei, respectively. The EGFP vector was used as a positive control. ( C ) Comparison of the number of EGFP-positive cells per well expressing either hGH::EGFP protein or EGFP alone ( N = 4). ( D ) Detection of hGH mRNA by semi RT-qPCR (upper) and RT-qPCR (lower) ( N = 3). Labels in X-axis show the expression vector with the presence (+) or absence (–) of the transfection reagent. ( E ) Western blotting for detection of EGFP, hGH::EGFP, and hGH::V5/His proteins using anti-EGFP antibody or anti-hGH antibodies. EGFP produced from E.coli was used as a positive control. ( F ) Western blotting of hGH::V5/His protein under reducing and non-reducing conditions using an anti-V5 antibody. Plus (+) and minus (–) represent treatment of protein samples with a reducing agent or PBS, respectively. ( G ) Quantity of hGH protein from hemocyte lysate measured by ELISA ( N = 5). ( H ) Schematic diagrams of two HiBiT-tagged hEPO expression vectors designed for scallop hemocytes (left) and HEK293 cells (right). ( I ) Relative quantification of hEPO secreted into the culture medium from scallop hemocytes and HEK293 cells using the HiBiT assay. ( J ) Western-blot analysis of hEPO expressed in scallop hemocytes and HEK293, with PNGase F treatment. Plus (+) and minus (–) represent treatment of protein samples with PNGase F or PBS, respectively

Article Snippet: hGH protein was quantified using a Human Growth Hormone ELISA Kit (R & D systems, USA) following the manufacture’s protocol.

Techniques: Expressing, Plasmid Preparation, Fluorescence, Imaging, Cell Culture, Staining, Positive Control, Comparison, Quantitative RT-PCR, Transfection, Western Blot, Produced, Enzyme-linked Immunosorbent Assay, Quantitative Proteomics