• Asian Pacific Journal of Cancer Prevention
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• v.16 no.12
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• pp.5115-5118
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• 2015

Background: The $p16^{INK4a}$ is a protein that expressed in Liquid-based cervical cytology specimens and has been proved link to cervical cancer. The $p16^{INK4a}$ could be detection by piezoelectric immunosensor and the immobilization of the $p16^{INK4a}$ antibody influence the sensitivity of the piezoelectric immunosensor. Materials and Methods: $5{\mu}L$ mouse polyclonal antibody against $p16^{INK4a}$ was bound onto the surface of immonosensor through two methods. (directly immobilized method; protein A method). Absorb of the $p16^{INK4a}$ antibody on the surface of immonosensor caused a shift in the resonant frequency of the immunosensor and The frequency changes recorded showed a better reproducibility. The activity of the immobilization antibody with the directly method and protein A method was tested with $p16^{INK4a}$ antigen. Results: The resonant frequency for different antibody immobilization methods were different, and the sensitivity for $p16^{INK4a}$ detection also different. Conclusions: The protein A method was found to be much more better than the directly method for the immobilization of the p16INK4A antibody on the gold electrode of the quartz crystal for cervical lesion detection. The Protein A method created more reproducible and stable immobilization antibody layers with p16INK4A antigen.

• Journal of Marine Bioscience and Biotechnology
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• v.6 no.1
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• pp.1-7
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• 2014

Marine-derived pathogens threat health and life of human and animals. Therefore, rapid and specific detection methods need to be developed. Here, we summarized various groups of detection methods, including conventional method, flow cytometry, nucleic acid-based method, and protein-based method. In addition, perspective of detection technique was discussed as a unified detection system for pathogens.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.153-157
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• 2017

This study was conducted to investigate the optimal artificial insemination (AI) time with diagnostic kit at ovulation time. We already applied the patent about the protein in the cow heat mucose in external reproductive tract. And we would examine the accuracy for detection of cow heat by the kit produced with the protein. Evaluation of optimal heat detection was tried two time at 12 hrs and 24 hrs after the heat. And then, AI service also performed two times with no relation to the results of heat diagnosis by heat detection kit and pregnancy rates were checked with rectal palpation on $60^{th}$ day after AI. Heat diagnostic results by kit in natural heat after 12 hrs in Hanwoo cows were showed 31.3~75.0% on positive in first heat detection and 33.3~100.0% on positve in second heat detection. In the $1^{st}$ positive results were significant different (p<0.05), but $2^{nd}$ positive were not. The results of heat detection showed different result on regional influence and individual cow effects. The pregnancy rates of first trial of heat detection were showed 34.4~78.7% on positive and 21.3~68.8% on negative after the diagnosis by heat detection kit. And the pregnancy rates of next trial of heat detection were showed 33.3~85.7% on positive and 14.3~66.6% on negative after the heat diagnosis. Both positive results of first trial and next trial also were showed significant different (p<0.05), but negative results were not. In positive result, first trial of total pregnancy rates was higher than the next trial of pregnancy, but there showed opposite results on negative results. In conclusion, the optimal heat detection kit is suitable to ordinary Hanwoo cows and it suggested that we have to improve the kit's accuracy by detecting the materials like proteins related optimal AI time.

• Animal Bioscience
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• v.36 no.6
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• pp.973-979
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• 2023

Objective: The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system, which is the most efficient and reliable tool for precisely targeted modification of the genome of living cells, has generated considerable excitement for industrial applications as well as scientific research. In this study, we developed a gene-editing and detection system for chick embryo sexing during the embryonic stage. Methods: By combining the CRISPR/Cas9 technical platform and germ cell-mediated germline transmission, we not only generated Z chromosome-targeted knockin chickens but also developed a detection system for fluorescence-positive male chicks in the embryonic stage. Results: We targeted a green fluorescent protein (GFP) transgene into a specific locus on the Z chromosome of chicken primordial germ cells (PGCs), resulting in the production of Z^GFP-knockin chickens. By mating Z^GFP-knockin females (Z^GFP/W) with wild males (Z/Z) and using a GFP detection system, we could identify chick sex, as the GFP transgene was expressed on the Z chromosome only in male offspring (Z^GFP/Z) even before hatching. Conclusion: Our results demonstrate that the CRISPR/Cas9 technical platform with chicken PGCs facilitates the production of specific genome-edited chickens for basic research as well as practical applications.

• Parasites, Hosts and Diseases
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• v.53 no.1
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• pp.85-93
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• 2015

Proteomic tools allow large-scale, high-throughput analyses for the detection, identification, and functional investigation of proteome. For detection of antigens from Haemaphysalis longicornis, 1-dimensional electrophoresis (1-DE) quantitative immunoblotting technique combined with 2-dimensional electrophoresis (2-DE) immunoblotting was used for whole body proteins from unfed and partially fed female ticks. Reactivity bands and 2-DE immunoblotting were performed following 2-DE electrophoresis to identify protein spots. The proteome of the partially fed female had a larger number of lower molecular weight proteins than that of the unfed female tick. The total number of detected spots was 818 for unfed and 670 for partially fed female ticks. The 2-DE immunoblotting identified 10 antigenic spots from unfed females and 8 antigenic spots from partially fed females. Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF) of relevant spots identified calreticulin, putative secreted WC salivary protein, and a conserved hypothetical protein from the National Center for Biotechnology Information and Swiss Prot protein sequence databases. These findings indicate that most of the whole body components of these ticks are non-immunogenic. The data reported here will provide guidance in the identification of antigenic proteins to prevent infestation and diseases transmitted by H. longicornis.

• Korean Journal of Veterinary Research
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• v.29 no.4
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• pp.541-548
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• 1989

The present study was carried out to determine viral antigens and its morphogenesis in the ultrathin frozen and araldite sections of cell cultures infected with ADV by protein A-gold labeling. ADV antigens were labeled with 10nm gold probes, and electron-dense gold particles were mainly present on viral nucleocapsids and viral envelopes. Immunogold labeling in the ultracryosections showed a very low degree of interaction with tissue structures. Immunogold labeling in the ultrathin cryosections can be useful tool for the detection of ADV antigens, and the technique also may provide its great potential for immunocytochemical studies on various virus-host cell Interactions.

• KSBB Journal
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• v.8 no.2
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• pp.143-149
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• 1993

Activated carbons were used to examine their performance for the separation of undesirable colored materials from culture broth containing hyaluronic acid. Six local samples and a NORIT ROX 08 were tested, whereas the latter was mainly studied under batch and continuous modes. The optimal wavelength for the detection of colored materials was 330nm. The optimal choice of NORIT ROX 08 provided 30% colored residuals with 96% hyaluronic acid recovery of original broth in batch experiments. The nonlinear adsorption behavior of protein and colored materials with activated carbon (C) was correlated by a Langmuir equation to give 18C/24+C and 500C/892+C for protein and colored materials, respectively. It appears that colored materials were composed of 78% protein and 22% glucose residuals on the basis of clearance results. A microscopic study using a scanning electron microscope suggests that regeneration of used activated carbon with 0.1N NaOH and hot water was not satisfactory. The present study proposes that the continuous monitoring of colored materials during purification can be accomplished by Installation of a UV monitor commonly used for continuous detection of protein during the process, as resulted from the significant correlation of color (A330)=0.353protein(mg/ml)+0.1(R=99.7%).

• Journal of fish pathology
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• v.27 no.2
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• pp.99-105
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• 2014

Biosensors consist of biochemical recognition agents like antibodies immobilized on the surfaces of transducers that change the recognition into a measurable electronic signal. Here we report a piezoelectric immunosensor made to detect Vibrio vulnificus. A 9MHz AT-cut piezoelectric wafer attached with two gold electrodes of 5mm diameter was used as the transducer of the QCM biosensor with a reproducibility of ${\pm}0.1Hz$ in frequency response. We have tried different approaches to immobilize antibody on the sensor chip. Concerning the orientation of antibody for the best antigen binding capacity, the antibody was immobilized by specific binding to protein G or by cross-linking through hydrazine. In addition, protein G was cross-linked on glutaraldehyde activated immine layer (PEI) or EDC/NHS activated sulfide monolayer (MPA). PEI was found to be more effective to immobilize protein G following glutaraldehyde activation than MPA. However, hydrazine chip showed a better capability to immobilize more IgG than protein G chip and a higher sensitivity. The sensor system was able to detect V. vulnificus in dose dependent manner and was able to detect bacterial cells within 5 minutes by monitoring frequency shifts in real time. The detection limit can be improved by preincubation to enrich the bacterial cell number.

• The Plant Pathology Journal
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• v.19 no.3
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• pp.159-161
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• 2003

The coat protein (CP) gene of Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was selected for the design of virus-specific primers for amplification and molecular detection of the virus in cultivated apple. A combined assay of reverse transcription and polymerase chain reaction (RT-PCR) was performed with a single pair of ApMV-specific primers and crude nucleic acid extracts from virus-infected apple for rapid detection of the virus. The PCR product was verified by restriction mapping analysis and by sequence determination. The lowest concentration of template viral RNA required for detection was 100 fg. This indicates that the RT-PCR for detection of the virus is a 10$^3$times more sensitive, reproducible and time-saving method than the enzyme-linked immunosorbent assay. The specificity of the primers was verified using other unrelated viral RNAs. No PCR product was observed when Cucumber mosaic virus (Cucumovirus) or a crude extract of healthy apple was used as a template in RT-PCR with the same primers. The PCR product (669 bp) of the CP gene of the virus was cloned into the plasmid vector and result-ant recombinant (pAPCP1) was selected for molecule of apple transformation to breed virus-resistant transgenic apple plants as the next step. This method can be useful for early stage screening of in vitro plantlet and genetic resources of resistant cultivar of apple plants.

• Proceedings of the Korean Vacuum Society Conference
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• 2013.08a
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• pp.268.2-268.2
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• 2013

Nanogap interdigitated electrodes (NIDEs) can serve as an alternative platform for the biomolecular detection [1]. In this work, the NIDEs were adopted in a simple and sensitive detection of Pneumococcal surface protein A (PspA). The NIDEs were fabricated by the combination of photo and chemical lithography. Photolithographically-defined initial gap of about 200 nm was narrowed down to a few tens of nanometers by surface-initiated growth of the initial electrodes (chemical lithography) [2]. Bare silicon oxide surface between the electrodes was chemically modified to immobilize capturing antibodies and, after exposure to the samples, the device was immersed in a solution containing the probe-antibody-conjugated Au nanoparticles (Au NPs). The conductance change accompanied with the Au NP immobilization was interpreted as the existence of PspA. Detection limit of the measurements and further improvement of the detection efficiency were discussed with the results from I-V analysis, scanning electron microscopy, and atomic force microscopy.