An immunomagnetic bead enrichment technique to improve the detection efficiency for trace silk protein, its application

Zheng, Hailing; Zhang, Wei; Yang, Hailiang; Ma, Chao; Zhou, Yang; Dai, Xianjun

doi:10.1016/j.culher.2019.02.003

Cited by 9 publications

(7 citation statements)

References 24 publications

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Section: Methodsmentioning

confidence: 99%

“…Silk protein monoclonal antibodies were diluted with coating buffer in different proportions, the SF-1, SF-2, and SF-5 antibodies were diluted at 1:2000, and the SF-3 antibody was diluted at 1:4000. 15 Fifty microliters of the silk protein monoclonal antibody solution was added to each well. For the blank control group, 50 μL of coating buffer was directly added to each well, the microtiter plate was coated at 4 °C overnight, and 300 μL of PBS was subsequently added to each well for washing, which was repeated three times.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

“…Immunomagnetic beads (IMBs) which can enrich and purify silk protein in solution were prepared by coupling silk protein antibody with magnetic beads. Then, the indirect enzyme-linked immunosorbent assay (ELISA) was used to evaluate the enrichment effect of the IMBs . This process involves the elution of silk protein on the IMBs and presents the problems of insufficient elution and complicated operations.…”

Section: Introductionmentioning

confidence: 99%

“…Then, the indirect enzymelinked immunosorbent assay (ELISA) was used to evaluate the enrichment effect of the IMBs. 15 This process involves the elution of silk protein on the IMBs and presents the problems of insufficient elution and complicated operations. For samples with trace amounts of protein, such as soil from archaeological sites, if the elution is insufficient, the test results will be affected.…”

Section: ■ Introductionmentioning

confidence: 99%

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Rapid Enrichment and Detection of Silk Residues from Tombs by Double-Antibody Sandwich ELISA Based on Immunomagnetic Beads

et al. 2021

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The extraction and identification of silk residues in tombs is of great significance for studying the distribution and spread of early silk. However, the complex organic matter in the tomb hinders the accurate identification of silk. In this study, a doubleantibody sandwich enzyme-linked immunosorbent assay (ELISA) based on immunomagnetic beads (IMBs) was developed for the rapid enrichment and detection of silk residues. The doubleantibody sandwich ELISA method established by pairing the IMBs prepared by the silk fibroin monoclonal antibody SF-3 and the silk fibroin monoclonal-labeled antibody bio-SF-1 had the highest detection sensitivity, with a linear detection range of 10 to 10 4 ng mL −1 and a detection limit of 5.12 ng mL −1 . This method was excellent in the extraction and analysis of silk residues from archaeological imprints and soil samples and successfully identified silk residues in samples at the final stage of silk degradation (physical invisible silk). The proteomics analysis results demonstrated the feasibility and practicability of this method.

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Section: Methodsmentioning

confidence: 99%

Section: ■ Materials and Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Rapid Enrichment and Detection of Silk Residues from Tombs by Double-Antibody Sandwich ELISA Based on Immunomagnetic Beads

et al. 2021

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“…Antibodies are proteins that are produced to fight against foreign molecules in the body but antibodies can also be used for molecule specific binding. Immunomagnetic beads have been used in applications including protein purification [18] and immunoassays [19]. There have also been several studies conducted using IMB isolation in forensic science applications.…”

Section: Immunomagnetic Beadsmentioning

confidence: 99%

Immunomagnetic Beads Coupled with Anti-PH-20 Antibodies to Isolate Sperm from DNA Mixtures for Sexual Assault Kit Application

Becks¹

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Sexual Assault Kits (SAKs) account for a large portion of DNA mixture samples that are processed within a forensic laboratory, most commonly in the form of vaginal swabs. A typical vaginal swab from a SAK contains the female victim's epithelial cells and varying concentrations of sperm cells left by the perpetrator. The key to analyzing these types of samples is to separate the mixture into male and female fractions. As such, there is a demand to develop novel techniques that have the capability of recovering as much of the male fraction as possible, specifically in cases where there is a limited sperm concentration to begin with. Magnetic beads are a versatile product in that they can be coupled with a variety of substances including a silica coating or various antibodies. Several studies have explored techniques to target testis specific molecules to isolate sperm from DNA mixtures. This study targeted a protein (PH-20) that is not testis specific but is intrinsic to the head of a sperm cell. The goal was to evaluate the efficiency of magnetic beads coupled with anti-PH-20 antibodies in isolating sperm cells from DNA mixtures. Results suggested that streptavidin (SA) coated immunomagnetic beads (IMBs) recover more male DNA than carboxylic acid coated IMBs in both male-only samples and male:female mixture samples. Overall, the SA-IMBs recovered approximately 21.7% of the total male DNA available in samples. While more research is needed to optimize the recovery of male DNA utilizing the IMB process, magnetic beads coupled with PH-20 antibodies have shown success in recovering male DNA and have considerable potential for forensic applications.

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On-chip immunomagnetic bead swarm based on magnetic actuation and mechanical vibration for biological detection

Pan

Gong

Saeed

et al. 2022

Sci. China Technol. Sci.

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An immunomagnetic bead enrichment technique to improve the detection efficiency for trace silk protein, its application

Cited by 9 publications

References 24 publications

Rapid Enrichment and Detection of Silk Residues from Tombs by Double-Antibody Sandwich ELISA Based on Immunomagnetic Beads

Rapid Enrichment and Detection of Silk Residues from Tombs by Double-Antibody Sandwich ELISA Based on Immunomagnetic Beads

Immunomagnetic Beads Coupled with Anti-PH-20 Antibodies to Isolate Sperm from DNA Mixtures for Sexual Assault Kit Application

On-chip immunomagnetic bead swarm based on magnetic actuation and mechanical vibration for biological detection

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