Stabilization of bio-sensors for in vivo applications
Inventors
Herbrechtsmeier, Peter • Müller, Achim • Knuth, Monika • Nikolaus, Katharina
Assignees
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Publication Number
US-8945942-B2
Publication Date
2015-02-03
Expiration Date
Abstract
The present invention relates to the use of preparations for stabilizing isolated proteins. In particular, the use of such a preparation for stabilizing receptors in biochemical sensors is disclosed. The invention in addition relates to biochemical sensors containing such preparations.
Core Innovation
The invention stabilizes isolated proteins, in particular receptor proteins used in biochemical sensors, by contacting the isolated protein with a cell lysate fraction that includes precanavalin. The cell lysate fraction is produced by lysing cells and separating off concanavalin A to obtain a cell lysate fraction comprising precanavalin for maintaining the function of concanavalin A.
The disclosure emphasizes that lysate fractions prepared from Canavalia ensiformis provide improved stabilization of concanavalin A, including recovery of function after exposure conditions such as autoclaving without loss of function. It also describes enriching or purifying precanavalin from a non-binding fraction and using the resulting preparation for maintaining concanavalin A function in a biochemical sensor context.
Biochemical sensor formulations are described in which concanavalin A is used together with precanavalin-containing cell lysate fraction. The sensor preparation includes co-formulation with other components such as dextran in hydrogel particles, including alginate beads, to maintain functionality and signal retention over time, as reflected by in vitro and longer-term stability comparisons.
Claims Coverage
The document provides two independent claims. They cover maintaining concanavalin A function in a biochemical sensor by contacting concanavalin A with a precanavalin-containing cell lysate fraction produced by separating concanavalin A, and a biochemical sensor that includes concanavalin A binding with high affinity to a substance to be detected together with a precanavalin-containing cell lysate fraction in contact with concanavalin A. A total of two dependent claims are specified here, focusing on sensor formulation and additional sensor context.
Contacting Concanavalin A with a Precanavalin-Containing Cell Lysate Fraction to Maintain Function
Maintaining the function of concanavalin A in a biochemical sensor by contacting concanavalin A with a cell lysate fraction comprising precanavalin.
Producing the Precanavalin-Containing Cell Lysate Fraction by Separating Concanavalin A
Producing the cell lysate fraction comprising precanavalin by producing the cell lysate from cells and separating the concanavalin A from the cell lysate to obtain the cell lysate fraction comprising precanavalin for maintaining the function of concanavalin A.
Stabilizing Concanavalin A Compared to Known Stabilizers
Using the cell lysate fraction comprising precanavalin as compared to known stabilizers to improve the stability of the concanavalin A.
Biochemical Sensor with High-Affinity Concanavalin A Binding and Precanavalin Lysate in Contact
Providing a biochemical sensor comprising concanavalin A that binds with high affinity to a substance that is to be detected and a cell lysate fraction comprising precanavalin, where the cell lysate fraction comprising precanavalin is in contact with concanavalin A to maintain the function of concanavalin A.
Dextran and Hydrogel Particles Co-Containing Concanavalin A and Precanavalin Lysate
Configuring the biochemical sensor additionally comprising dextran and concanavalin A in hydrogel particles.
Overall, the claims cover sensor systems in which concanavalin A function is maintained by contacting it with a cell lysate fraction comprising precanavalin produced by separating concanavalin A, with asserted stabilization improvement relative to known stabilizers. The sensor is further defined by high-affinity binding to a detected substance and, in dependent form, by inclusion of dextran and placement of concanavalin A together with the precanavalin-containing lysate fraction in hydrogel particles.
Stated Advantages
Improves the stability of concanavalin A as compared to known stabilizers.
Maintains the function of concanavalin A in the biochemical sensor.
Documented Applications
Biochemical sensors that use concanavalin A, together with a cell lysate fraction comprising precanavalin, including co-formulation with dextran in hydrogel particles (e.g., alginate beads) for glucose sensor stability comparisons.
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