On chip real time monitoring of B-cells hybridoma secretion of immunoglobulin.
The secretions of molecules by cells are of tremendous interest for both fundamental insights studies and medical purposes. In this study, we propose a new biochip-based approach for the instantaneous monitoring of protein secretions, using antibody production by B lymphocytes cultured in vitro. This was possible thanks to the Surface Plasmon Resonance imaging (SPRi) of a protein biochip where antigen proteins (Hen Egg Lysozyme, HEL) were micro-arrayed along with series of control proteins. B cell hybridomas were cultured on the chip and the secretion of immunoglobulins (antibody) specific to HEL was monitored in real-time and detected within only few minutes rather than after a 30-60 min incubation with standard ELISA experiments. This fast and sensitive detection was possible thanks to the sedimentation of the cells on the biochip sensitive surface, where local antibody concentrations are much higher before dilution in the bulk medium. An other interesting feature of this approach for the secretion monitoring was the independence of the SPR response--after normalization--regarding to the density of the surface-immobilized probes. Such biosensor might thus pave the way to new tools capable of both qualitative and semi-quantitative analysis of proteins secreted by other immune cells.
CREAB Group, SPRAM Laboratory UMR 5819 (CEA-CNRS-UJF), Institute for Nanosciences and Cryogeny, CEA-Grenoble, 38054 Grenoble, France.
Sarah
S
Milgram
Sandra
S
Cortes
Marie-Bernadette
MB
Villiers
Patrice
P
Marche
Arnaud
A
Buhot
Thierry
T
Livache
Yoann
Y
Roupioz
D001402:B-Lymphocytes / Q000378:metabolism*; D015374:Biosensing Techniques / Q000295:instrumentation*; D018929:Cell Culture Techniques / Q000295:instrumentation; D002460:Cell Line; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D006825:Hybridomas / Q000378:metabolism*; D007118:Immunoassay / Q000295:instrumentation*; D007136:Immunoglobulins / Q000096:biosynthesis*; D040081:Protein Array Analysis / Q000295:instrumentation*; D020349:Surface Plasmon Resonance / Q000295:instrumentation*
D007136:Immunoglobulins