Feasibility of singlicate-based analysis in bridging ADA assay on Meso-Scale Discovery platform: comparison with duplicate analysis

Bioanalysis ◽  
2021 ◽  
Author(s):  
Zhihua Jiang ◽  
John Kamerud ◽  
Zhiping You ◽  
Soma Basak ◽  
Elena Seletskaia ◽  
...  
Keyword(s):  
Ligand Binding ◽  
Coefficient Of Variation ◽  
Performance Characteristics ◽  
Supportive Evidence ◽  
Binding Assays ◽  
Meso Scale Discovery ◽  
Minimal Impact ◽  
Platform Comparison ◽  
The Impact ◽  
Meso Scale

Aim: To investigate the feasibility of singlicate analysis in anti-drug antibody (ADA) assay by comparing performance characteristics for assays qualified in duplicate and singlicate formats. Materials & methods: We employed modeling to assess and quantify the impact of singlicate to cut point factor (CPF) in scenarios with the duplicate precision from 1–20% and the proportion of well-to-well variance to overall assay variance from 0.01–0.90. The impact to CPF by singlicate is marginal if the well-to-well coefficient of variation is <10% and represents <25% of the overall variability. Results & conclusion: The assay parameters including sensitivity, precision, selectivity, drug and target tolerance were comparable between singlicate and duplicate based assays. Our results suggested the minimal impact of singlicate analysis on ADA assay with good duplicate precision. The study provided additional supportive evidence that the singlicate-based analysis is feasible in ADA ligand binding assays.

scholarly journals Application of multiplexed pharmacokinetic immunoassay to quantify in vivo drug forms and coadministered biologics

Bioanalysis ◽  
2019 ◽  
Vol 11 (24) ◽  
pp. 2251-2268 ◽  
Author(s):  
Nicole Woodbury ◽  
Eric Bald ◽  
Brian Geist ◽  
Tong-Yuan Yang
Keyword(s):  
Clinical Study ◽  
Ligand Binding ◽  
Plasma Concentrations ◽  
Serum Concentrations ◽  
Binding Assays ◽  
Meso Scale Discovery ◽  
Methods Development ◽  
Nonclinical Study ◽  
Meso Scale

Aim: Meso Scale Discovery U-PLEX® provides an opportunity to develop multiplexed pharmacokinetic (PK) immunoassays. Two case studies demonstrate the utility of multiplexed PK methods. Materials & methods: Development of PK ligand-binding assays quantify of nonclinical plasma concentrations of a biotherapeutic that has degraded due to in vivo biotransformation, and clinical serum concentrations from two biotherapeutics spiked into a single sample. Results: Data from multiplexed U-PLEX PK methods are comparable to results from single-readout streptavidin Meso Scale Discovery gold PK methods. Multiplex measurement of a nonclinical study showed acceptable performance for accuracy, precision and dilutional linearity while a clinical study additionally passed selectivity, specificity and stability. Conclusion: Regulated, validation-ready multiplex PK methods for both nonclinical and clinical studies allow opportunities for high-throughput bioanalysis.

Recommendations for singlet-based approach in ligand binding assays: an IQ Consortium perspective

Bioanalysis ◽  
2020 ◽  
Author(s):  
Renuka Pillutla ◽  
Boris Gorovits ◽  
Carol Gleason ◽  
Jorge Quiroz ◽  
David Christopher ◽  
...  
Keyword(s):  
Ligand Binding ◽  
Simulation Models ◽  
Study Quality ◽  
Binding Assays ◽  
Pharmaceutical Development ◽  
Drug Candidates ◽  
Leadership Group ◽  
The Impact ◽  
Analytical Platforms ◽  
Good Agreement

Historically, ligand-binding assays for pharmacokinetic samples employed duplicate rather than singlet-based analysis. Herein, the Translational and absorption, distribution, metabolism and excretion (ADME) Sciences Leadership Group of the International Consortium for Innovation and Quality in Pharmaceutical Development (IQ) presents a study aiming to determine the value of duplicate versus singlet-based testing. Based on analysis of data collected from eight organizations for 20 drug candidates representing seven molecular types and four analytical platforms, statistical comparisons of validation and in-study quality controls and study unknown samples demonstrated good agreement across duplicate sets. Simulation models were also used to assess the impact of sample duplicate characteristics on bioequivalence outcomes. Results show that testing in singlet is acceptable for assays with % CV ≤15% between duplicates. Singlet-based approach is proposed as the default for ligand-binding assays while a duplicate-based approach is needed where imprecision and/or inaccuracy impede the validation of the assay.

A competitive ligand-binding assay to detect neutralizing antibodies to a bispecific drug using a used multiplex Meso Scale Discovery platform

Bioanalysis ◽  
2021 ◽  
Author(s):  
Vitaly Ablamunits ◽  
Soma Basak ◽  
Rosemary Lawrence-Henderson ◽  
Teresa M Caiazzo ◽  
John Kamerud
Keyword(s):  
Ligand Binding ◽  
Neutralizing Antibodies ◽  
Binding Assay ◽  
Large Molecule ◽  
Ligand Binding Assay ◽  
Meso Scale Discovery ◽  
Candidate Drug ◽  
Meso Scale ◽  
Competitive Ligand ◽  
Better Than

Background: Monitoring appearance of neutralizing antibodies (NAbs) to multidomain large molecule drugs is a challenging task. Materials & methods: Here, we report development of a competitive ligand-binding assay for detection of NAbs to a bispecific candidate drug using a used multiplex Meso Scale Discovery platform, which allows for detection of NAbs to both drug arms in the same sample. Results: The assay has sensitivity better than 250 ng/ml and is tolerant to the presence of drug at concentration >600 μg/ml and to the level of soluble target(s) >400 ng/ml. Conclusion: Our data suggest that multiplex approach can be successfully used for development of NAb assays in competitive ligand-binding assay format.

scholarly journals Structural basis for Na+-sensitivity in dopamine D2 and D3 receptors

2015 ◽  
Vol 51 (41) ◽  
pp. 8618-8621 ◽  
Author(s):  
Mayako Michino ◽  
R. Benjamin Free ◽  
Trevor B. Doyle ◽  
David R. Sibley ◽  
Lei Shi
Keyword(s):  
Ligand Binding ◽  
Binding Site ◽  
Computational Analysis ◽  
Structural Basis ◽  
Ligand Binding Site ◽  
Binding Assays ◽  
D3 Receptors ◽  
Dopamine D2 ◽  
The Impact

To understand the structural basis for the Na+-sensitivity of ligand binding to dopamine D2-like receptors, using computational analysis in combination with binding assays, we identified interactions critical in propagating the impact of Na+on receptor conformations and on the ligand-binding site.

Development of an Meso Scale Discovery ligand-binding assay for measurement of free (drug-unbound) target in nonhuman primate serum

Bioanalysis ◽  
2021 ◽  
Author(s):  
Yun Liu ◽  
Ronald Robinson ◽  
Thao Ung ◽  
Chrysanthe Spais ◽  
Justin Schreiber ◽  
...  
Keyword(s):  
Ligand Binding ◽  
Nonhuman Primate ◽  
Binding Assay ◽  
Free Form ◽  
Ligand Binding Assay ◽  
Target Engagement ◽  
Meso Scale Discovery ◽  
Drug Interference ◽  
Assay Performance ◽  
Meso Scale

Aim: To quantify the free form of a protein as a target-engagement biomarker in nonhuman primate serum, a Meso Scale Discovery ligand-binding assay was developed and qualified. Results: The initial assay produced an unexpected artifact when used to measure the free target in study samples dosed with drug. By using incurred study samples dosed with high drug levels to test assay performance, we developed an alternative assay that does not suffer from drug interference. Conclusion: Our work demonstrated that an assay designed to measure free target may not necessarily deliver reliable quantitation. In our case, incurred study samples dosed with drug proved to be useful in developing an alternative free assay that does not suffer from drug interference.

Adventures in critical reagent lot changes in ligand-binding assays: redevelopment, bridging and additional processing requirements

Bioanalysis ◽  
2021 ◽  
Author(s):  
Alissa Rauwerdink ◽  
Michael Benson ◽  
Allison Jayne ◽  
Sathyapriya Babu ◽  
Jessica St Charles ◽  
...  
Keyword(s):  
Ligand Binding ◽  
Method Development ◽  
Close Monitoring ◽  
Binding Assays ◽  
Entire Study ◽  
First Case ◽  
Assay Performance ◽  
And Performance ◽  
The Impact

Aim: Critical reagents have significant impact on ligand-binding assay performance. The critical reagents selected during method development should be well-evaluated, as the quality of these reagents will dictate performance of the assay over time. Critical reagents in ligand-binding assays are almost always produced using a biological system, so batch yield, purity and performance tend to vary greatly. Due to the essential nature of critical reagents in the assay, changes in critical reagents can have dramatic impact on the assay and results, so close monitoring of assay performance is required. Methodology & results: We present here three examples of critical reagent lot changes that required creative solutions to maintain assay performance. The first case study is an example of the impact of different lots of analyte within a quantitative assay that resulted in the need to redevelop the assay in a different format. Case study two outlines an assay where a surrogate matrix is the critical reagent in an assay and the difficulties encountered over the course of several years and lot changes. The third case study covers an immunogenicity assay with a commercial detection that did not have sufficient quantity to cover the entire study lifecycle. As a result of the reagent change, a new assay was developed. Discussion & conclusion: A robust plan for critical reagent generation and lifecycle management should be adapted in order to avoid costly delays and rework. The performance of an assay depends on the continuity of the critical reagent supply. Reagents should be carefully selected to include the binding and performance properties required for an assay.

scholarly journals Critical reagents for ligand-binding assays: process development methodologies to enable high-quality reagents

Bioanalysis ◽  
2022 ◽  
Author(s):  
Caroline Kittinger ◽  
Jared Delmar ◽  
Lisa Hewitt ◽  
Rebecca Holcomb ◽  
Christopher Jones ◽  
...  
Keyword(s):  
Ligand Binding ◽  
Process Development ◽  
Process Conditions ◽  
Generation Process ◽  
Binding Assays ◽  
Performance Control ◽  
Development Approach ◽  
Immunogenicity Assays ◽  
The Impact

Development of biotherapeutics require pharmacokinetic/pharmacodynamic (PK/PD) and immunogenicity assays that are frequently in a ligand-binding assay (LBA) format. Conjugated critical reagents for LBAs are generated conjugation of the biotherapeutic drug or anti-drug molecule with a label. Since conjugated critical reagent quality impacts LBA performance, control of the generation process is essential. Our perspective is that process development methodologies should be integrated into critical reagent production to understand the impact of conjugation reactions, purification techniques and formulation conditions on the quality of the reagent. In this article, case studies highlight our approach to developing process conditions for different molecular classes of critical reagents including antibodies and a peptide. This development approach can be applied to the generation of future conjugated critical reagents.

Challenges and Issues in the Design of Micro-Machined Antennas - A Review

2020 ◽  
Vol 13 ◽  
Author(s):  
Ashish Kumar ◽  
Amar Partap Singh Pharwaha
Keyword(s):  
Patch Antenna ◽  
Ground Plane ◽  
Substrate Material ◽  
Review Article ◽  
High Index ◽  
Performance Characteristics ◽  
Machining Process ◽  
Patch Antennas ◽  
Micro Machining ◽  
The Impact

Background: Patch antennas are composed of the substrate material with patch and ground plane on the both sides of the substrate. The dimensions and performance characteristics of the antenna are highly influenced by the choice of the appropriate substrate depending upon the value of their dielectric constant. Generally, low index substrate materials are used to design the patch antenna but there are also some of the applications, which require the implementation of patch antenna design on high index substrate like silicon and gallium arsenide. Objective: The objective of this article is to review the design of antennas developed on high index substrate and the problems associated with the use of these materials as substrate. Also, main challenges and solutions have been discussed to improve the performance characteristics while using the high index substrates. Method: The review article has divided into various sections including the solution of the problems associated with the high index substrates in the form of micro-machining process. Along with this, types of micro machining and their applications have discussed in detail. Results: This review article investigates the various patch antennas designed with micro-machining technology and also discusses the impact of micro-machining process on the performance parameters of the patch antennas designed on high index substrates. Conclusion: By using the micro-machining process, the performance of patch antenna improves drastically but fabrication and tolerances at such minute structures is very tedious task for the antenna designers.

scholarly journals The structure of SeviL, a GM1b/asialo-GM1 binding R-type lectin from the mussel Mytilisepta virgata

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Kenichi Kamata ◽  
Kenji Mizutani ◽  
Katsuya Takahashi ◽  
Roberta Marchetti ◽  
Alba Silipo ◽  
...  
Keyword(s):  
Immune System ◽  
Sialic Acid ◽  
Ligand Binding ◽  
Steric Hindrance ◽  
Sequence Similarity ◽  
Binding Assays ◽  
Subunit Interface ◽  
Nmr Techniques ◽  
And Control ◽  
Asialo Gm1

AbstractSeviL is a recently isolated lectin found to bind to the linear saccharides of the ganglioside GM1b (Neu5Ac$$\alpha$$ α (2-3)Gal$$\beta$$ β (1-3)GalNAc$$\beta$$ β (1-4)Gal$$\beta$$ β (1-4)Glc) and its precursor, asialo-GM1 (Gal$$\beta$$ β (1-3)GalNAc$$\beta$$ β (1-4)Gal$$\beta$$ β (1-4)Glc). The crystal structures of recombinant SeviL have been determined in the presence and absence of ligand. The protein belongs to the $$\beta$$ β -trefoil family, but shows only weak sequence similarity to known structures. SeviL forms a dimer in solution, with one binding site per subunit, close to the subunit interface. Molecular details of glycan recognition by SeviL in solution were analysed by ligand- and protein-based NMR techniques as well as ligand binding assays. SeviL shows no interaction with GM1 due to steric hindrance with the sialic acid branch that is absent from GM1b. This unusual specificity makes SeviL of great interest for the detection and control of certain cancer cells, and cells of the immune system, that display asialo-GM1.

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