ecoHSA™ Animal-free Human Serum Albumin

• Improves health of cells by acting as a fatty acid carrier and hydrophobic moiety scavenger.
• Stabilizes membrane proteins and maintains oncotic pressure for better cultures and results.
• Extends life of cell cultures by scavenging toxins.
• Promotes cell growth by reducing or eliminating the use of FBS in media.
• Minimizes variability between experiments because of batch-to-batch consistency.
• Eliminates risk of animal or viral infectious agents in cell cultures.
• Helps comply with FDA recommendations for animal-free components.
• Provides high quality at a value price.

ecoHSA™ Animal-free Human Serum Albumin is an animal-free preparation of the human albumin gene produced in the seeds of Asian Rice (Oryza sativa). Recent advances in seed protein production technologies have turned rice endosperm into an optimal system for expressing recombinant proteins.

ecoHSA™ white paper

Description	Datasheet	Qty	Order
ecoHSA™ Animal Free Human Serum Albumin		2 x 0.5 gm
ecoHSA™ Animal Free Human Serum Albumin		10 gm

Albumins have multiple uses and are of profound value in cell culture work. They are often used in the biomanufacture of therapeutic monoclonal antibodies and recombinant proteins, and are an important component in serum-free cell culture. In particular, Human Serum Albumin (HSA) is typically used as a supplement. HSA is added to tissue culture media because it is thought to maintain the stability of cell membranes and chelate trace amounts of toxic components present in media and cell culture containers. Additionally, albumins have been effectively used for cell washing and cryopreservation to name a few applications.

In the past, all commercial human albumin proteins were extracted from plasma. However, existing and new regulations are discouraging the use of animal components, even in early research studies. Additionally, human or animal derived albumin proteins, even those produced in vitro using insect or mammalian cultures, suffer from variable performance due to low yields and/or production complexities. The need for a defined, consistent and animal-free HSA is more important than ever.

ecoHSA™ is an animal-free preparation of the human albumin gene produced in the seeds of Asian Rice (Oryza sativa). Recent advances in seed protein production technologies have turned rice endosperm into an optimal system for expressing recombinant proteins. Furthermore, the manufacturing process of ecoHSA™ uses rigorous standards and procedures, so the end product is of the highest performance and quality possible.

A primary function of ecoHSA™ is to stabilize a range of important small molecules and ions. It acts as an antioxidant with multiple functions, such as binding and protection of fatty acids, sequestering copper and keeping it from oxidative reactions, and protecting amino acids and other molecules from oxidation. ecoHSA™ can be used in multiple areas of research, such as: development of mammalian cell cultures, cell storage and cryopreservation, coating for cell culture vessels and devices, formulation of potential protein therapeutics (in vitro testing), testing of vaccine formulations and manufacturing, infertility treatment research, molecular delivery, in vitro diagnostics and more.

Testing And R&D ecoHSA™ Animal Free Human Serum Albumin has been extensively tested in multiple functional studies.It has been proven to be similar in size and function to native HSA in all tests performed. Therefore, ecoHSA™ can become a suitable, and even better substitute, than HSA that is extracted from traditional animal or human sources.

Figure 1 below shows a Coomassie blue stained gel (left) and a silver nitrate stained gel (right) comparing size and content of competitive HSA products. The figure shows that there is no difference in migration patterns or purity between: 1) ecoHSA™ , 2) a commercial human plasma serum albumin purchased as a lyophilized powder (pHSA-P, 96-99% pure), or 3) a commercial human plasma serum albumin purchased as a liquid (pHSA-P, >96% pure).

Figure 2 below shows that ecoHSA™ has a purity greater than 95%, as measured on an Agilent 1200, with an HPLC-SEC protein column 5A, and detected at wavelength = 220 nm (DRD).

Figure 3 below shows the relative activity of CHO (Chinese Hamster Ovary) cells grown in the absence or presence of ecoHSA™ or pHSA-P (see Section B for explanation), and under different concentrations. Cells were cultured 48 hours under each concentration of ecoHSA™ and pHSA-P in D-MEM/F12 media. Cell activity was determined by an MTT (Methyl Thiazolyl Tetrazolium) assay. This data shows that ecoHSA™ has the same or better effect on cell activity as the most popular commercially available HSA product.

Figure 4 below shows CHO cell viability when grown at different concentrations of ecoHSA™ over time. Cells were continuously cultured in D-MEM/F12 containing 0.0, 0.5, 1.0, and 2.0 g/L of ecoHSA™ . Methyl Thiazolyl Tetrazolium assays were performed at 24, 72, 120, and 168 hours to determine cell activity and health. The amount of 1.0 g/L ecoHSA™ (violet line) is the most suitable and recommended for overall cell culture conditions.

CHO cells were incubated in D-MEM/F12 media with no additional supplement, or supplemented with 2 g/L ecoHSA™ , 2 g/ L pHSA-P, 10% FBS, or no protein additive for 36 hours. Cells were then collected and suspended in 70% ethanol at 4C overnight. Fixed cells were centrifuged at 2000 X g for 5 minutes and washed with ice-cold PBS. Cells were incubated with 100 mg/ml RNaseA in PBS for 30min at 37C then stained in the dark with PI (20 mg/ml) for 30 min at 4C. A minimum of 10,000 cells were counted and analyzed by flow cytometry. This figure shows that the effect of ecoHSA™ on the cell cycle is the same as pHSA-P, especially when comparing G1 and S phase percentages between the two proteins.

ecoHSA™ is an animal-free recombinant HSA produced in rice endosperm. It has been shown to be structurally and biochemically equivalent to plasma-derived human serum albumin, but without any of the side effects and potential problems associated with human or animal derived HSA or cell culture additives like FBS. ecoHSA™ will not only help reduce or eliminate the use of fetal bovine serum in media, but it will also increase cell survival, promote cell growth, and improve your research results. ecoHSA™ has all the benefits of native HSA in cell culture, yet provides better consistency, safety, and regulatory compliance.

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