Biochip-based instruments development for space exploration: influence of the antibody immobilization process on the biochip resistance to freeze-drying, temperature shifts and cosmic radiations

Abstract

International audienceDue to the diversity of antibody (Ab)-based biochips chemistries available and the little knowledge about biochips resistance to space constraints, immobilization of Abs on the surface of the biochips dedicated to Solar System exploration is challenging. In the present paper, we have developed ten different biochip models including covalent or affinity immobilization with full-length Abs or Ab fragments. Ab immobilizations were carried out in oriented/non-oriented manner using commercial activated surfaces with N-hydroxysuccinic ester (NHS-surfaces) or homemade surfaces using three generations of dendrimers (dendrigraft of poly L-lysine (DGL) surfaces). The performances of the Ab -based surfaces were cross-compared on the following criteria: (i) analytical performances (expressed by both the surface density of immobilized Abs and the amount of antigens initially captured by the surface) and (ii) resistance of surfaces to preparation procedure (freeze-drying, storage) or spatial constraints (irradiation and temperature shifts) encountered during a space mission. The latter results have been expressed as percentage of surface binding capacity losses (or percentage of remaining active Abs). The highest amount of captured antigen was achieved with Ab surfaces having full-length Abs and DGL-surfaces that have much higher surface densities than commercial NHS-surface. After freeze-drying process, thermal shift and storage sample exposition, we found that more than 80% of surface binding sites remained active in this case. In addition, the resistance of Ab surfaces to irradiation with particles such as electron, carbon ions or protons depends not only on the chemistries (covalent/affinity linkages) and strategies (oriented/non-oriented) used to construct the biochip, but also on the type, energy and fluence of incident particles. Our results clearly indicate that full-length Ab immobilization on NHS-surfaces and DGL-surfaces should be preferred for potential use in instruments for planetary exploration

Everything You Always Wanted to Know about Poly‐l‐lysine Dendrigrafts (But Were Afraid to Ask)

“…Indeed, it was demonstrated that DGLs are readily soluble in water at least up to 60 gL À1 in water, [12] are stable under sterilization conditions, [28] and are partially degradable under the action of endogenous peptidases. Indeed, it was demonstrated that DGLs are readily soluble in water at least up to 60 gL À1 in water, [12] are stable under sterilization conditions, [28] and are partially degradable under the action of endogenous peptidases.…”

“…In order to enter the real world of industrial applications,s uch as drug and gene delivery,b iomaterial and tissue engineering, bioimaging or biosensing, polymers should display an umber of crucial propertiest hat we believe DGLs exhibit. Indeed, it was demonstrated that DGLs are readily soluble in water at least up to 60 gL À1 in water, [12] are stable under sterilization conditions, [28] and are partially degradable under the action of endogenous peptidases. [29] In addition, these biomacromolecules are non-immunogenic [30] and carry low cytotoxicity (i.e., IC50 values on Hep2 cellso f1 6.8, 13.2, and 13.9 mgmL À1 from second-to fourth-generation DGL, respectively), [31] in comparison with other polycationic polymers (e.g.,a bout 90 %v iability for DGL G3 versusl ess than 50 %v iability for hyperbranched PEI at as ame concentration of 12.5 mgmL À1 on HeLa cells).…”

The non-destructive separation of diverse astrobiologically relevant organic molecules by customizable capillary zone electrophoresis and monolithic capillary electrochromatography

“…Thus, the very polymers that are indicative of life are rendered undetectable with GC-MS (Poinot and Geffroy-Rodier, 2015). Other analytical strategies proposed for space missions include biosensors (Coussot et al ., 2017) and antibody-based systems such as the Life Marker Chip (Rix et al ., 2011). However, these approaches lack flexibility as the instruments must be built with specific molecular targets.…”

Photochemistry on the Space Station—Aptamer Resistance to Space Conditions: Particles Exposure from Irradiation Facilities and Real Exposure Outside the International Space Station

“…One main concern relies on the resistance of biochips to the cumulative effects of these radiations. In recent years, studies have been performed to evaluate the resistance of antibody reagents to space constraints, especially regarding the effect of specific particles on antibody binding performances (Le Postollec et al, 2009a;Le Postollec et al, 2009b;Baqué et al, 2011a;de Diego-Castilla et al, 2011;Baqué et al, 2017;. To complement ground-based radiation studies, a short term mission was performed on the BIOPAN-6 low-earth orbit platform to demonstrate the effects of cumulative radiations on the antibodies' ability to bind to their respective antigens (Derveni et al, 2012;Derveni et al, 2013).…”