EcoHSA Animal Free Human Serum Albumin Campaign

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1 EcoHSA Animal Free Human Serum Albumin Campaign GENERAL INFORMATION Albumins have multiple uses and are of profound value in multiple laboratory experiments, especially in cell culture work. Genlantis ecohsa is produced using a novel technology platform that produces a highly pure and active recombinant HSA( see Figure 1). In the following experiments, we demonstrate that ecohsa performs as well as animal- or plasma-derived HSA, on par with supplementation, but without the dangerous or detrimental contamination threats that may come from animal-derived HSA. Expression Vector EXPERIMENTAL METHOD CHO-K1, Vero and Hybridoma cells were each cultured in 96 well plates using the appropriate cell culture media with or without additional additives. The cells were resuspended in the appropriate medium and cultured for 10 days continuously. Cell viability and cell density were tested daily, and the antibody secretion level was measured when the cells reached 90% confluency. Methyl Thiazolyl Tetrazolium (MTT) assay was used to determine cell activity at 24, 72, 120, and 168 hours. Genetic Transformation Plant Regeneration RESULTS 1. ecohsa performance in CHO-K1 Cell Culture CHO-K1 Cell Viability 5% FBS 5% FBS eco + ecohsa + phsa Endosperm Expression Scale up & Processing Formulation & End Product Figure 1: ecohsa Technology Platform.

2 CHO-K1 Cell Density eco + ecohsa 5% FBS + phsa Cell viability (MTT) and density tests show that when CHO-K1 culture media is supplemented with ecohsa, the amount of FBS can be reduced by half (from 10% to 5%) without noticeable negative effect on either cell viability or density. Furthermore, experiments show that there is no difference in the viability or density of cells with plasma-derived HSA (phsa) versus ecohsa when using the same amount of FBS in the media. 2. ecohsa performance in Vero Cell Culture Vero Cell Viability + ecohsa + phsa

3 Vero Cell Density 5% FBS + ecohsa 5% FBS + phsa MTT cell viability tests, as well as density measurements on cultured Vero cells show that ecohsa supplementation of media can help reduce the amount of FBS used while keeping results near the levels obtained with only. 3. ecohsa performance in Hybridoma SP2/O Cell Culture SP2/O Cell Viability 5% FBS + ecohsa + phsa

4 SP2/O Cell Density 5% FBS 5% FBS+ecoHSA + phsa In hybridoma cells, both cell viability and density reached the highest levels when cells were cultured in media supplemented with either or with + ecohsa, with no difference seen between plasma HSA (phsa) and ecohsa Antibody Efficiency P/N phsa + ecohsa Antibody production test using ELISA in SP2/O cells show that these hybridomas produced the best antibody secretion results when cultured in media supplemented with either or with media containing + ecohsa. In this test, the ecohsa performance exceeded that of plasma HSA (phsa) by nearly 50% when used in media containing.

5 SUMMARY AND CONCLUSION Our cell proliferation assay results clearly show that ecohsa performed the same or better than plasma HSA (phsa) when used to supplement cell culture media in CHO-K1, Vero, and Hybridoma cells. When used along with in culture media, ecohsa was as effective in maintaining high cell viability and density as supplementation alone. We clearly demonstrate that ecohsa can be safely and reliably used as an alternative to plasma HSA (phsa) in serum-free or serum reduced cell culture media while eliminating any safety concerns of animal- or human-derived products. ecohsa Animal Free Human Serum Albumin Size Catalog # 2 x 0.5 gm H gm H