Thermally denatured BSA, a surrogate additive to replace BSA in buffers for high-throughput screening.
The use of thermally denatured bovine serum albumin (tdBSA) as an additive in high-throughput screening (HTS) buffers has been studied with the aim of finding a surrogate to native albumin devoid of its inconveniences, in particular its compound masking effect. The presence of aggregates in the thermally denatured material did not have any negative impact on common readout technologies used in HTS such as fluorescence intensity (FLINT), fluorescence polarization, time-resolved fluorescence resonance energy transfer (TR-FRET) and luminescence. tdBSA rendered the same beneficial effects as native albumin in several assays or even improved its performance due to the lack of specific binding properties. Although tdBSA still binds compounds nonspecifically as any other protein does, it mitigates the compound masking effect observed with native albumin and can be postulated as a convenient surrogate to BSA for HTS purposes.
Peña
Imanol
I
Biological Reagents and Assay Development Department, GlaxoSmithKline, Madrid, Spain.
Domínguez
Juan Manuel
JM
D000818:Animals; D002021:Buffers; D002417:Cattle; D057075:Enzyme Assays / Q000379:methods*; D005454:Fluorescence Polarization; D031541:Fluorescence Resonance Energy Transfer; D057166:High-Throughput Screening Assays / Q000379:methods*; D008163:Luminescent Measurements; D011489:Protein Denaturation; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000378:metabolism
D002021:Buffers; D012710:Serum Albumin, Bovine