| Title |
A rapid solution-based method for determining the affinity of heroin hapten-induced antibodies to heroin, its metabolites, and other opioids
|
|---|---|
| Published in |
Analytical & Bioanalytical Chemistry, April 2018
|
| DOI | 10.1007/s00216-018-1060-4 |
| Pubmed ID | |
| Authors |
Oscar B. Torres, Alexander J. Duval, Agnieszka Sulima, Joshua F. G. Antoline, Arthur E. Jacobson, Kenner C. Rice, Carl R. Alving, Gary R. Matyas |
| Abstract |
We describe for the first time a method that utilizes microscale thermophoresis (MST) technology to determine polyclonal antibody affinities to small molecules. Using a novel type of heterologous MST, we have accurately measured a solution-based binding affinity of serum antibodies to heroin which was previously impossible with other currently available methods. Moreover, this mismatch approach (i.e., using a cross-reactive hapten tracer) has never been reported in the literature. When compared with equilibrium dialysis combined with ultra-performance liquid chromatography/tandem mass spectrometry (ED-UPLC/MS/MS), this novel MST method yields similar binding affinity values for polyclonal antibodies to the major heroin metabolites 6-AM and morphine. Additionally, we herein report the method of synthesis of this novel cross-reactive hapten, MorHap-acetamide-a useful analog for the study of heroin hapten-antibody interactions. Using heterologous MST, we were able to determine the affinities, down to nanomolar accuracies, of polyclonal antibodies to various abused opioids. While optimizing this method, we further discovered that heroin is protected from serum esterase degradation by the presence of these antibodies in a concentration-dependent manner. Lastly, using affinity data for a number of structurally different opioids, we were able to dissect the moieties that are crucial to antibody binding. The novel MST method that is presented herein can be extended to the analysis of any ligand that is prone to degradation and can be applied not only to the development of vaccines to substances of abuse but also to the analysis of small molecule/protein interactions in the presence of serum. Graphical abstract Strategy for the determination of hapten-induced antibody affinities using Microscale thermophoresis. |
X Demographics
The data shown below were collected from the profiles of 6 X users who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 2 | 33% |
| Mexico | 1 | 17% |
| Germany | 1 | 17% |
| Unknown | 2 | 33% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 3 | 50% |
| Members of the public | 2 | 33% |
| Science communicators (journalists, bloggers, editors) | 1 | 17% |
Mendeley readers
The data shown below were compiled from readership statistics for 21 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 21 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 6 | 29% |
| Student > Ph. D. Student | 3 | 14% |
| Student > Bachelor | 2 | 10% |
| Professor | 1 | 5% |
| Other | 1 | 5% |
| Other | 2 | 10% |
| Unknown | 6 | 29% |
| Readers by discipline | Count | As % |
|---|---|---|
| Pharmacology, Toxicology and Pharmaceutical Science | 3 | 14% |
| Chemistry | 3 | 14% |
| Biochemistry, Genetics and Molecular Biology | 2 | 10% |
| Medicine and Dentistry | 2 | 10% |
| Agricultural and Biological Sciences | 1 | 5% |
| Other | 1 | 5% |
| Unknown | 9 | 43% |
Attention Score in Context
This research output has an Altmetric Attention Score of 3. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 25 May 2018.
All research outputs
#15,151,839
of 25,461,852 outputs
Outputs from Analytical & Bioanalytical Chemistry
#4,563
of 9,646 outputs
Outputs of similar age
#179,893
of 341,133 outputs
Outputs of similar age from Analytical & Bioanalytical Chemistry
#55
of 173 outputs
Altmetric has tracked 25,461,852 research outputs across all sources so far. This one is in the 40th percentile – i.e., 40% of other outputs scored the same or lower than it.
So far Altmetric has tracked 9,646 research outputs from this source. They receive a mean Attention Score of 3.1. This one has gotten more attention than average, scoring higher than 52% of its peers.
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We're also able to compare this research output to 173 others from the same source and published within six weeks on either side of this one. This one has gotten more attention than average, scoring higher than 66% of its contemporaries.