Quantitative assessment of bovine serum albumin proteins for blocking applications

Abstract

Bovine serum albumin (BSA) is one of the most widely used protein reagents in the scientific community, especially for surface passivation ("blocking") applications in various bioassays. Numerous BSA protein options are commercially available, however, there is scarce information about which ones are preferable for blocking applications.Herein, we conducted biophysical and bioassay measurements to quantitatively compare the conformational, adsorption, and blocking properties of BSA protein reagents that were obtained through six purification methods. Depending on the method, there were significant differences in the conformational and adsorption properties of BSA proteins, mainly due to the presence of fatty acid stabilizers. In turn, we discovered that fatty acidfree BSA proteins exhibit superior blocking performance to fatty acid-stabilized BSA proteins in surface-and nanoparticle-based bioassays. We critically discuss mechanistic factors behind these performance variations and our findings offer a practical framework to guide BSA selection for blocking applications.A common element of bioassays is the need to minimize nonspecific binding of biomolecules in order to maximize assay specificity and sensitivity 1, 2 . To meet this objective, a wide range of reagents, ranging from crude materials such as milk proteins to synthetic nanomaterials such as finely tuned polymers, have been developed to coat target surfaces and to minimize nonspecific binding events-a function that is commonly referred to as

A matrigel-free method to generate matured human cerebral organoids using 3D-Printed microwell arrays

“…Passivation coatings generally render a PDMS surface hydrophilic, preventing protein adsorption and cell-adhesion [ 53 ]. For BSA, albumin protein adsorbs to the PDMS surface like any protein, which renders the surface hydrophilic [ 49 , 54 , 55 ], creating an anti-fouling layer that inhibits other protein adsorption. However, protein-based anti-fouling coatings are temporary because the PDMS regains its hydrophobicity over time.…”

“…Besides, uncoated devices had less than half EB formation efficiency compared to the most efficient conditions (mPEG and Lipidure). Additionally, the prevalent formation of multiple EBs in BSA coated devices is likely due to the stability of BSA coating, which is vulnerable to culture conditions and is likely to crack into multiple small coated areas [ 54 , 55 ]. Furthermore, device curvature did not seem to salvage BSA coated devices, as BSA-coated HRC (curved) and HRF (flat) devices showed an insignificant difference in EB formation efficiency.…”

Development of a Cryoprotective Formula Based on the Fucose-Containing Polysaccharide FucoPol

“…11,12,19 BSA and Euro-Collins solutions presented a Newtonian behavior with lower viscosity (Figure 1). This fact is related to the globular structure of BSA molecules 20 and to the low-molecular-weight components of the Euro-Collins solution, which disable strong entanglements and friction between molecules, resulting in lower apparent viscosity values. The FP + BSA solution (Figure 1, black-blue) presented an apparent viscosity and a shear-thinning behavior similar to the FucoPol solution (FP).…”

On-The-Spot Sampling and Detection of Viral Particles on Solid Surfaces Using a Sponge Virus Sensor Incorporated with Finger-Press Fluid Release

“…This characteristic is crucial because hydrophilic surfaces are inherently less susceptible to biofouling compared to hydrophobic ones due to the competitive binding of water molecules which reduces the chances of foulant attachment. 50,51 Additionally, BSA was utilized as a blocking agent to create a physical barrier against the nonspecific binding of proteins 52,53 to the surface of the BSA/antibody/rGO/graphene/ sponge electrode.…”