Articles and methods involving detection of binding
Inventors
Scholle, Michael D. • SHERRILL, John
Assignees
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Publication Number
US-12135330-B2
Publication Date
2024-11-05
Expiration Date
Abstract
Articles and methods involving detection of binding between two or more species using MALDI are generally provided. In some embodiments, a method comprises exposing a first species to a composition comprising a second species, capturing the first species on a self-assembled monolayer, and performing MALDI on the self-assembled monolayer to generate a signal from the second species and a signal from a standard species present in the self-assembled monolayer. A ratio (first ratio) of the signal from the second species to the signal from the standard species may be compared to a cutoff value in order to determine if binding has occurred. This method may allow determination of a relative affinity of the second species for the first species.
Core Innovation
The invention relates to a method for detecting binding between a first species and a second species using MALDI with a self-assembled monolayer that comprises a standard species. The first species is exposed to a composition comprising the second species and then captured on the self-assembled monolayer, and MALDI is performed to generate a signal from the second species and a signal from the standard species.
Binding is determined by comparing the signal from the second species to the signal from the standard species. The method uses a ratio of the signal from the second species to the signal from the standard species, and determines binding has occurred when the ratio is greater than or equal to a cutoff value.
The workflow is framed around reproducible quantitative or semi-quantitative MALDI enabled by signal normalization to the standard species signal, and around quality-control abnormality detection using the standard or control signals. Example implementations in the document include SAMDI for small-molecule-protein binding and pooled screening.
Claims Coverage
The patent provides two independent methods for detecting binding. Across these claims, the inventive features center on MALDI signal generation for a second species and a standard species from a self-assembled monolayer, and determining binding by a cutoff based on a signal ratio. The two independent claims differ mainly in how the self-assembled monolayer and standard species are defined.
MALDI binding detection with standard species on self-assembled monolayer
A method for detecting binding between a first species and a second species by exposing the first species to a composition comprising the second species, capturing the first species on a self-assembled monolayer comprising a standard species prior to the capture of the first species thereon, performing MALDI on the self-assembled monolayer to generate a signal from the second species and a signal from the standard species, measuring both signals, and determining whether binding has occurred if a ratio of the signal from the second species to the signal from the standard species is greater than or equal to a cutoff value.
Standard species with bonding or interacting group to support on self-assembled monolayer
A method for detecting binding between a first species and a second species by exposing the first species to a composition comprising the second species, capturing the first species on a self-assembled monolayer comprising a standard species where the standard species comprises a group capable of bonding and/or interacting with a support on which the self-assembled monolayer is disposed and/or that is bonded to and/or interacting with the support on which the self-assembled monolayer is disposed, performing MALDI to generate a signal from the second species and a signal from the standard species, measuring both signals, and determining whether binding has occurred if a ratio of the signal from the second species to the signal from the standard species is greater than or equal to a cutoff value.
Both independent claims define binding detection using MALDI signals from the second species and a standard species on a self-assembled monolayer, with binding called by whether a normalized second/standard signal ratio meets or exceeds a cutoff value. One independent claim requires the standard species to be present on the self-assembled monolayer prior to capturing the first species, while the other further specifies that the standard species includes a group capable of bonding and/or interacting with a support.
Stated Advantages
Provides reproducible quantitative or semi-quantitative MALDI via standard-signal normalization.
Enables determining binding using a normalized signal ratio (second/standard) compared to a cutoff value.
Supports quality control or abnormality detection using the standard or control signals.
Allows SAMDI workflow for small-molecule-protein binding, including pooled screening.
Documented Applications
SAMDI workflow for small-molecule-protein binding, including pooled compound binder identification.
Application to binding detection with example binders including β-secretase (BACE) and dual leucine zipper kinase (DLK).
Pooled screening for binder identification using the disclosed MALDI-based standard-normalized signal ratio approach.
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