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.

The effectiveness of quality control samples in pharmaceutical bioanalysis

Bioanalysis ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 135-145
Author(s):  
Stephen Keller ◽  
Jorge Quiroz ◽  
Dave Christopher ◽  
Enaksha Wickremsinhe ◽  
Wanping Geng ◽  
...  
Keyword(s):  
Quality Control ◽  
Systematic Evaluation ◽  
Pharmacokinetic Data ◽  
Concentration Data ◽  
Pharmaceutical Development ◽  
Regulatory Guidance ◽  
Leadership Group ◽  
Continued Use ◽  
Analytical Platforms

The use of quality control (QC) samples in bioanalysis is well established and consistent with regulatory guidance. However, a systematic evaluation of whether QC samples serve the intended purpose of improving data quality has not been undertaken. The Translational and ADME Sciences Leadership Group (TALG) of the International Consortium for Innovation and Quality in Pharmaceutical Development (IQ) conducted an evaluation to assess whether closer agreement is observed when comparing pharmacokinetic data from two passed runs, than when comparing data from failed and passed (retest) runs. Analysis of data collected across organizations, molecular types and analytical platforms, revealed that bioanalytical methods are very reproducible; and that QC samples improve the overall quality of pharmacokinetic concentration data and justifies their continued use.

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.

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.

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.

Modeling of Tread Block Contact Mechanics Using Linear Viscoelastic Theory3

2008 ◽  
Vol 36 (3) ◽  
pp. 211-226 ◽  
Author(s):  
F. Liu ◽  
M. P. F. Sutcliffe ◽  
W. R. Graham
Keyword(s):  
High Speed ◽  
Slip Rate ◽  
Material Model ◽  
Contact Forces ◽  
Block Modeling ◽  
Rate Dependent ◽  
Linear Viscoelastic Material ◽  
Linear Viscoelastic ◽  
The Impact ◽  
Good Agreement

Abstract In an effort to understand the dynamic hub forces on road vehicles, an advanced free-rolling tire-model is being developed in which the tread blocks and tire belt are modeled separately. This paper presents the interim results for the tread block modeling. The finite element code ABAQUS/Explicit is used to predict the contact forces on the tread blocks based on a linear viscoelastic material model. Special attention is paid to investigating the forces on the tread blocks during the impact and release motions. A pressure and slip-rate-dependent frictional law is applied in the analysis. A simplified numerical model is also proposed where the tread blocks are discretized into linear viscoelastic spring elements. The results from both models are validated via experiments in a high-speed rolling test rig and found to be in good agreement.

Dynamic model for high-speed rotors based on their experimental characterization

2017 ◽  
Vol 2 (4) ◽  
pp. 25
Author(s):  
L. A. Montoya ◽  
E. E. Rodríguez ◽  
H. J. Zúñiga ◽  
I. Mejía
Keyword(s):  
Dynamic Model ◽  
High Speed ◽  
Natural Frequencies ◽  
Mode Shapes ◽  
Experimental Characterization ◽  
Rotating Systems ◽  
Computing Performance ◽  
The Impact ◽  
Stiffness Distribution ◽  
Good Agreement

Rotating systems components such as rotors, have dynamic characteristics that are of great importance to understand because they may cause failure of turbomachinery. Therefore, it is required to study a dynamic model to predict some vibration characteristics, in this case, the natural frequencies and mode shapes (both of free vibration) of a centrifugal compressor shaft. The peculiarity of the dynamic model proposed is that using frequency and displacements values obtained experimentally, it is possible to calculate the mass and stiffness distribution of the shaft, and then use these values to estimate the theoretical modal parameters. The natural frequencies and mode shapes of the shaft were obtained with experimental modal analysis by using the impact test. The results predicted by the model are in good agreement with the experimental test. The model is also flexible with other geometries and has a great time and computing performance, which can be evaluated with respect to other commercial software in the future.

scholarly journals Measuring Hydrometeors Using a Precipitation Microphysical Characteristics Sensor: Sampling Effect of Different Bin Sizes on Drop Size Distribution Parameters

2018 ◽  
Vol 2018 ◽  
pp. 1-15 ◽  
Author(s):  
Xichuan Liu ◽  
Taichang Gao ◽  
Yuntao Hu ◽  
Xiaojian Shu
Keyword(s):  
Drop Size ◽  
Rain Rate ◽  
Monte Carlo Model ◽  
Rain Gauge ◽  
Sampling Schemes ◽  
Distribution Parameters ◽  
Optimum Sampling ◽  
Simulation Results ◽  
The Impact ◽  
Good Agreement

In order to improve the measurement of precipitation microphysical characteristics sensor (PMCS), the sampling process of raindrops by PMCS based on a particle-by-particle Monte-Carlo model was simulated to discuss the effect of different bin sizes on DSD measurement, and the optimum sampling bin sizes for PMCS were proposed based on the simulation results. The simulation results of five sampling schemes of bin sizes in four rain-rate categories show that the raw capture DSD has a significant fluctuation variation influenced by the capture probability, whereas the appropriate sampling bin size and width can reduce the impact of variation of raindrop number on DSD shape. A field measurement of a PMCS, an OTT PARSIVEL disdrometer, and a tipping bucket rain Gauge shows that the rain-rate and rainfall accumulations have good consistencies between PMCS, OTT, and Gauge; the DSD obtained by PMCS and OTT has a good agreement; the probability of N0, μ, and Λ shows that there is a good agreement between the Gamma parameters of PMCS and OTT; the fitted μ-Λ and Z-R relationship measured by PMCS is close to that measured by OTT, which validates the performance of PMCS on rain-rate, rainfall accumulation, and DSD related parameters.

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.

scholarly journals Responses of the Pheromone-Binding Protein of the Silk Moth Bombyx mori on a Graphene Biosensor Match Binding Constants in Solution

Sensors ◽  
2021 ◽  
Vol 21 (2) ◽  
pp. 499
Author(s):  
Caroline Bonazza ◽  
Jiao Zhu ◽  
Roger Hasler ◽  
Rosa Mastrogiacomo ◽  
Paolo Pelosi ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Binding Protein ◽  
Binding Constants ◽  
Isoamyl Acetate ◽  
The Other ◽  
Oxide Surface ◽  
Binding Assays ◽  
Pheromone Binding Protein ◽  
Silk Moth ◽  
Good Agreement

An electronic biosensor for odors was assembled by immobilizing the silk moth Bombyx mori pheromone binding protein (BmorPBP1) on a reduced graphene oxide surface of a field-effect transistor. At physiological pH, the sensor detects the B. mori pheromones, bombykol and bombykal, with good affinity and specificity. Among the other odorants tested, only eugenol elicited a strong signal, while terpenoids and other odorants (linalool, geraniol, isoamyl acetate, and 2-isobutyl-3-methoxypyrazine) produced only very weak responses. Parallel binding assays were performed with the same protein and the same ligands, using the common fluorescence approach adopted for similar proteins. The results are in good agreement with the sensor’s responses: bombykol and bombykal, together with eugenol, proved to be strong ligands, while the other compounds showed only poor affinity. When tested at pH 4, the protein failed to bind bombykol both in solution and when immobilized on the sensor. This result further indicates that the BmorPBP1 retains its full activity when immobilized on a surface, including the conformational change observed in acidic conditions. The good agreement between fluorescence assays and sensor responses suggests that ligand-binding assays in solution can be used to screen mutants of a binding protein when selecting the best form to be immobilized on a biosensor.

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