Layer-by-layer coated digitally encoded microcarriers for quantification of proteins in serum and plasma.
The "layer-by-layer" (LbL) technology has been widely investigated for the coating of flat substrates and capsules with polyelectrolytes. In this study, LbL polyelectrolyte coatings applied at the surface of digitally encoded microcarriers were evaluated for the quantitative, sensitive, and simultaneous detection of proteins in complex biological samples like serum, plasma, and blood. LbL coated microcarriers were therefore coupled to capture antibodies, which were used as capture agents for the detection of tumor necrosis factor (TNF-alpha), P24, and follicle stimulating hormone (FSH). It was found that the LbL coatings did not disassemble upon incubating the microcarriers in serum and plasma. Also, nonspecific binding of target analytes to the LbL coating was not observed. We showed that the LbL coated microcarriers can reproducibly detect TNF-alpha, P24, and FSH down to the picogram per milliliter level, not only in buffer but also in serum and plasma samples. Microcarrier-to-microcarrier intratube variations were less then 30%, and interassay variations less than 8% were observed. This paper also shows evidence that the LbL coated digitally encoded microcarriers are ideally suited for assaying proteins in "whole" blood in microfluidic chips, which are of high interest for "point-of-care" diagnostics.
Laboratory of General Biochemistry and Physical Pharmacy, University of Ghent, Harelbekestraat 72, 9000 Ghent, Belgium.
Stefaan
S
Derveaux
Barbara G
BG
Stubbe
Chris
C
Roelant
Marc
M
Leblans
Bruno G
BG
De Geest
Jo
J
Demeester
Stefaan C
SC
De Smedt
D000818:Animals; D000906:Antibodies / Q000276:immunology; D015374:Biosensing Techniques / Q000295:instrumentation*; D001798:Blood Proteins / Q000032:analysis*; D002021:Buffers; D002417:Cattle; D004573:Electrolytes / Q000737:chemistry; D005640:Follicle Stimulating Hormone / Q000032:analysis / Q000097:blood; D016655:HIV Core Protein p24 / Q000032:analysis / Q000097:blood; D006801:Humans; D008164:Luminescent Proteins / Q000737:chemistry; D051379:Mice; D008863:Microspheres; D010949:Plasma / Q000737:chemistry*; D011137:Polystyrenes / Q000737:chemistry*; D040081:Protein Array Analysis; D044967:Serum / Q000737:chemistry* / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000097:blood; D000097573:Red Fluorescent Protein
D000906:Antibodies; D001798:Blood Proteins; D002021:Buffers; D004573:Electrolytes; D005640:Follicle Stimulating Hormone; D016655:HIV Core Protein p24; D008164:Luminescent Proteins; D011137:Polystyrenes; D014409:Tumor Necrosis Factor-alpha; D000097573:Red Fluorescent Protein