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
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
Benefits And General Applications of ecoHSA
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.
Main characteristics of ecoHSA
The following table shows that ecoHSA has the same essential characteristics as native HSA.
Purity and Size of Competitive HSA Products by SDS-PAGE
|Amino acid sequence
Based on DNA sequence
Identical to Native HSA DNA sequence|
Molecular mass (MALDl) (66.5kD)
66.9 - 68.1 kDa
66.8 - 67.9 kDa|
Yes, not statistically different from native HSA|
|Tryptic peptide analysis
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).
Purity of ecoHSA
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).
Functional Testing of ecoHSA in CHO cells
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
CHO Cells Activity at Different Concentration of ecoHSA Over Time
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.
F-Cell Cycle Comparison Between ecoHSA and pHSA-P
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.