Two-peak approximation in kinetic capillary electrophoresis
Bibliographic record
Abstract
Kinetic capillary electrophoresis (KCE) constitutes a toolset of homogeneous kinetic affinity methods for measuring rate constants of formation (k(+)) and dissociation (k(-)) of non-covalent biomolecular complexes, C, formed from two binding partners, A and B. A parameter-based approach of extracting k(+) and k(-) from KCE electropherograms relies on a small number of experimental parameters found from the electropherograms and used in explicit expressions for k(+) and k(-) derived from approximate solutions to mass transfer equations. Deriving the explicit expressions for k(+) and k(-) is challenging but it is justified as the parameter-based approach is the simplest way of finding k(+) and k(-) from KCE electropherograms. Here, we introduce a unique approximate analytical solution of mass transfer equations in KCE termed a "two-peak approximation" and a corresponding parameter-based method for finding k(+) and k(-). The two-peak approximation is applicable to any KCE method in which: (i) A* binds B to form C* (the asterisk denotes a detectable label on A), (ii) two peaks can be identified in a KCE electropherogram and (iii) the concentration of B remains constant. The last condition holds if B is present in access to A* and C* throughout the capillary. In the two-peak approximation, the labeling of A serves only for detection of A and C and, therefore, is not required if A (and thus C) can be observed with a label-free detection technique. We studied the proposed two-peak approximation, in particular, its accuracy, by using the simulated propagation patterns built with the earlier-developed exact solution of the mass-transfer equations for A* and C*. Our results prove that the obtained approximate solution of mass transfer equations is correct. They also show that the two-peak approximation facilitates finding k(+) and k(-) with a relative error of less than 10% if two peaks can be identified on a KCE electropherogram. Importantly, the condition of constant concentration of B is always satisfied in macroscopic approach to studying kinetics at equilibrium (MASKE) whether or not B is in excess to A* and C*, and, thus, the two-peak approximation is applicable to MASKE. It completes a toolset of fitting-free methods for processing MASKE data and makes MASKE a simple practical method for finding k(+) and k(-) of "fast", "slow", and "intermediate-rate" non-covalent interactions.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.001 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".