Multiplexed suspension array platform for high-throughput protein assays

Abstract Background As a common small molecule substance, environmental hormones widely exist in nature, especially water sources, which have a profound effects in humans. Highly efficient and sensitive method for estrogens in the environment are essential. Results In this paper, a novel high-throughput platform was established based on five small hormones molecules specificity aptamer and magnetic beads (MBs). The results showed that the sensitivity of the proposed method are greatly improved. The limit of detection(LOD) of this method for atrazine(Atz), profenofos, bisphenolA(BPA), estradiol(E2), and polychlorinated biphenyls(PCBs) were 9.46, 20.75, 23.81, 8.97, 6.27 pg/mL, respectively. The Recovery rate of the diluted environmental hormones spiked in the samples of Haihe river were in the range of 87.5-111.02% with relative standard deviations (RSDs) lower than 28.44%. Conclusion This platform based on new complementary strand fragments can simultaneously rapid detection five environmental hormones. The whole procedure completed within 1.5h including sample treatment, incubation and detection, greatly improving the detection efficiency.

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Combination of Suspension Array and Mycelial Growth Assay for Detecting Multiple-Fungicide Resistance in Botrytis cinerea in Hebei Province in China

Plant Disease ◽

10.1094/pdis-07-18-1269-re ◽

2019 ◽

Vol 103 (6) ◽

pp. 1213-1219

Author(s):

Zehua Su ◽

Xin Zhang ◽

Jianjiang Zhao ◽

Wenqiao Wang ◽

Lei Shang ◽

...

Keyword(s):

Botrytis Cinerea ◽

High Throughput ◽

Fungicide Resistance ◽

Mycelial Growth ◽

Accurate Method ◽

Hebei Province ◽

Combination Method ◽

Major Type ◽

Suspension Array ◽

Growth Assay

To provide a high-throughput, efficient, and accurate method to monitor multiple-fungicide resistance of Botrytis cinerea in the field, we used the suspension array, sequencing, and mycelial growth assay in our research. Discriminating-dose bioassays for detecting carbendazim, diethofencarb, boscalid, and iprodione resistance (CarR, DieR, BosR, and IprR, respectively) were used to analyze 257 isolates collected from Hebei Province in China during 2016 and 2017. High resistance frequencies to carbendazim (100%), diethofencarb (92.08%), and iprodione (86.59%) were detected. BosR isolates accounted for 11.67% of the total. In addition, 103 isolates were randomly selected for phenotype and genotype detection. The high-throughput suspension array was utilized to detect eight genotypes simultaneously, including BenA-E198, BenA-198A, SdhB-H272, SdhB-272Y, BcOS1-I365, BcOS1-365S, erg27-F412, and erg27-412S, which were associated with resistance toward carbendazim or diethofencarb, boscalid, iprodione, and fenhexamid (FenR), respectively. Most of the benzimidazole-resistant isolates (81.55%) possessed the E198V mutation in the BenA gene. Ninety-three isolates with dual resistance to carbendazim and diethofencarb showed the E198V/K mutation. All BosR isolates carried the H272R mutation in the SdhB gene. The I365S and Q369P+N373S (66.99%) mutations in the BcOS1 gene were more frequently observed. No mutation was detected in the erg27 gene in Hebei isolates. There were 13 resistance profile phenotypes. Phenotypes with triple resistance were the most common (83.50%), and CarRDieRBosSIprRFenS was the major type. CarR isolates that carried E198V/K/A were all highly resistant (HR) and only one F200Y mutant was moderately resistant (MR) to carbendazim. Isolates that possessed E198V/K were MR or HR to diethofencarb. BosR isolates that possessed H272R mutation were lowly resistant (LR). IprR isolates were all LR or MR. The distribution of half maximal effective concentrations of CarR isolates with E198V/K mutations and IprR isolates with Q369P+N373S mutations significantly increased from 2016 to 2017. Combined with our observations, a combination method of the high-throughput suspension array and the mycelial growth assay was suggested to accurately monitor multiple resistance of B. cinerea in the field.

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High-Throughput 3D Tumor Spheroid Array Platform for Evaluating Sensitivity of Proton-Drug Combinations

International Journal of Molecular Sciences ◽

10.3390/ijms23020587 ◽

2022 ◽

Vol 23 (2) ◽

pp. 587

Author(s):

Dong Woo Lee ◽

Jung Eun Kim ◽

Ga-Haeng Lee ◽

Arang Son ◽

Hee Chul Park ◽

...

Keyword(s):

Cell Lines ◽

High Throughput ◽

High Throughput Screening ◽

Quantitative Data ◽

Survival Rates ◽

Drug Combinations ◽

Targeted Drugs ◽

Tumor Spheroid ◽

Array Platform ◽

Hnscc Cell

Proton beam therapy (PBT) is a critical treatment modality for head and neck squamous cell carcinoma (HNSCC). However, not much is known about drug combinations that may improve the efficacy of PBT. This study aimed to test the feasibility of a three-dimensional (3D) tumor-spheroid-based high-throughput screening platform that could assess cellular sensitivity against PBT. Spheroids of two HNSCC cell lines—Fadu and Cal27—cultured with a mixture of Matrigel were arrayed on a 384-pillar/well plate, followed by exposure to graded doses of protons or targeted drugs including olaparib at various concentrations. Calcein staining of HNSCC spheroids revealed a dose-dependent decrease in cell viability for proton irradiation or multiple targeted drugs, and provided quantitative data that discriminated the sensitivity between the two HNSCC cell lines. The combined effect of protons and olaparib was assessed by calculating the combination index from the survival rates of 4 × 4 matrices, showing that Cal27 spheroids had greater synergy with olaparib than Fadu spheroids. In contrast, adavosertib did not synergize with protons in both spheroids. Taken together, we demonstrated that the 3D pillar/well array platform was a useful tool that provided rapid, quantitative data for evaluating sensitivity to PBT and drug combinations. Our results further supported that administration of the combination of PBT and olaparib may be an effective treatment strategy for HNSCC patients.

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Novel Electrochemiluminescent Assays for Drug Discovery

JALA Journal of the Association for Laboratory Automation ◽

10.1016/s1535-5535-04-00049-8 ◽

2000 ◽

Vol 5 (1) ◽

pp. 45-48 ◽

Cited By ~ 2

Author(s):

Maura C. Kibbey ◽

David MacAllan ◽

James W. Karaszkiewicz

Keyword(s):

Drug Discovery ◽

High Precision ◽

High Throughput ◽

High Performance ◽

Dynamic Range ◽

Wide Dynamic Range ◽

Receptor Ligand ◽

Enzymatic Assays ◽

Detection Systems ◽

Protein Assays

IGEN's ORIGEN® technology, which is based on electrochemiluminescence, has been adopted by a number of research and bioanalytical laboratories who have recognized its exquisite sensitivity, high precision, wide dynamic range, and flexibility in formatting a wide variety of applications. IGEN's M-SERIES™ marks the introduction of the second generation of detection systems employing the ORIGEN technology specifically repackaged to address the needs of the high throughput laboratories involved in drug discovery. Assays are formatted without wash steps. Users realize the high performance of a heterogeneous technology with the convenience of a homogeneous format. The M-SERIES platform can address enzymatic assays (kinases, proteases, helicases, etc.), receptor-ligand or protein-protein assays, immunoassays, quantitation of nucleic acids, as well as other applications. Recent assay formats will be explored in detail.

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Simultaneous detection of five antibiotics in milk by high-throughput suspension array technology

Talanta ◽

10.1016/j.talanta.2011.05.040 ◽

2011 ◽

Vol 85 (2) ◽

pp. 1160-1165 ◽

Cited By ~ 36

Author(s):

Pu Su ◽

Nan Liu ◽

Maoxiang Zhu ◽

Baoan Ning ◽

Ming Liu ◽

...

Keyword(s):

High Throughput ◽

Simultaneous Detection ◽

Suspension Array ◽

Array Technology

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High-throughput bead based suspension array for the detection of acute respiratory viral pathogens among children aged <5 years in Pakistan

International Journal of Infectious Diseases ◽

10.1016/j.ijid.2016.11.253 ◽

2016 ◽

Vol 53 ◽

pp. 100-101

Author(s):

F. Aziz ◽

J. Samad ◽

I. Rizvi ◽

A. Sami ◽

S. Qureshi ◽

...

Keyword(s):

High Throughput ◽

Viral Pathogens ◽

Suspension Array

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Analytical Validation of the Tag-It High-Throughput Microsphere-Based Universal Array Genotyping Platform: Application to the Multiplex Detection of a Panel of Thrombophilia-Associated Single-Nucleotide Polymorphisms

Clinical Chemistry ◽

10.1373/clinchem.2004.035071 ◽

2004 ◽

Vol 50 (11) ◽

pp. 2028-2036 ◽

Cited By ~ 47

Author(s):

Susan Bortolin ◽

Margot Black ◽

Hemanshu Modi ◽

Ihor Boszko ◽

Daniel Kobler ◽

...

Keyword(s):

Single Nucleotide Polymorphisms ◽

High Throughput ◽

Nucleotide Polymorphisms ◽

Sequencing Data ◽

Single Nucleotide ◽

Direct Dna Sequencing ◽

Array Platform ◽

Genotyping Platform ◽

Universal Array ◽

Primer Sets

Abstract Background: We have developed a novel, microsphere-based universal array platform referred to as the Tag-It™ platform. This platform is suitable for high-throughput clinical genotyping applications and was used for multiplex analysis of a panel of thrombophilia-associated single-nucleotide polymorphisms (SNPs). Methods: Genomic DNA from 132 patients was amplified by multiplex PCR using 6 primer sets, followed by multiplex allele-specific primer extension using 12 universally tagged genotyping primers. The products were then sorted on the Tag-It array and detected by use of the Luminex xMAP™ system. Genotypes were also determined by sequencing. Results: Empirical validation of the universal array showed that the highest nonspecific signal was 3.7% of the specific signal. Patient genotypes showed 100% concordance with direct DNA sequencing data for 736 SNP determinations. Conclusions: The Tag-It microsphere-based universal array platform is a highly accurate, multiplexed, high-throughput SNP-detection platform.

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Dual-spectra encoded suspension array using reversed-phase microemulsion UV curing and electrostatic self-assembling

RSC Advances ◽

10.1039/c8ra02410c ◽

2018 ◽

Vol 8 (38) ◽

pp. 21272-21279 ◽

Cited By ~ 2

Author(s):

Guangxia Feng ◽

Qinghua He ◽

WenYue Xie ◽

Yonghong He ◽

Xuejing Chen ◽

...

Keyword(s):

High Throughput ◽

Rapid Growth ◽

Uv Curing ◽

Reversed Phase ◽

Suspension Array ◽

Self Assembling ◽

Modern Biotechnology

The rapid growth of demand for high-throughput multiplexed biochips from modern biotechnology has led to growing interest in suspension array based on multi-channel encoded microbeads.

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