HRP-conjugated-nanobody-based cELISA for rapid and sensitive clinical detection of ASFV antibodies was written by Zhao, Huijun;Ren, Jiahui;Wu, Shuya;Guo, Haoran;Du, Yongkun;Wan, Bo;Ji, Pengchao;Wu, Yanan;Zhuang, Guoqing;Zhang, Angke;Zhang, Gaiping. And the article was included in Applied Microbiology and Biotechnology in 2022.Synthetic Route of C9H18O5S The following contents are mentioned in the article:
African swine fever (ASF), which is caused by the ASF virus (ASFV), is a highly contagious hemorrhagic disease that causes high mortality to domestic porcine and wild boars and brings huge economic losses to world swine industry. Due to the lack of an effective vaccine, the control of ASF must depend on early, efficient, and cost-effective detection and strict control and elimination strategies. Traditional serol. testing methods are generally associated with high testing costs, complex operations, and high tech. requirements. As a promising alternative diagnostic tool to traditional antibodies, nanobodies (Nb) have the advantages of simpler and faster generation, good stability and solubility, and high affinity and specificity, although the system is dependent on the immunization of Bactrian camels to obtain the specific VHH library of the target protein. The application of Nbs in the detection of ASFV antibodies has not yet been reported yet. Using a phage display technol., one Nb against the ASFV p54 protein that exhibited high specificity and affinity, Nb8, was successfully screened. A HEK293T cell line stably expressing Nb8-horseradish peroxidase (HRP) fusion protein was established using the lentiviral expression system. Following the optimization of the reaction conditions, the Nb8-HRP fusion protein was successfully used to establish a competitive ELISA (cELISA) to detect ASFV-specific antibodies in pig serum, for the first time. There was no cross-reaction with healthy pig serum, porcine pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), porcine epidemic diarrhea virus (PEDV), and classical swine fever virus (CSFV) pos. sera. The optimal cut-off value for the cELISA by ROC anal. was 52.5%. A total of 209 serum samples were tested using the developed cELISA and a com. ELISA kit. The results showed that the relative specificity of the cELISA was 98.97%, and the relative sensitivity of the cELISA was 93.3%, with the percent agreement between the two ELISA methods being 98.56%. In conclusion, a specific, sensitive, and repeatable cELISA was successfully developed based on the Nb8 as a probe, providing a promising method for the detection of anti-ASFV antibodies in clin. pig serum. We successfully screened a specific, high affinity nanobody against ASFV p54 protein. We establish a method for continuous and stable expression of Nb-HRP fusion protein using a lentiviral packaging system. We establish a nanobody cELISA detection method that can monitor an ASF infection. This study involved multiple reactions and reactants, such as (2R,3R,4S,5R,6S)-2-(Hydroxymethyl)-6-(isopropylthio)tetrahydro-2H-pyran-3,4,5-triol (cas: 367-93-1Synthetic Route of C9H18O5S).
(2R,3R,4S,5R,6S)-2-(Hydroxymethyl)-6-(isopropylthio)tetrahydro-2H-pyran-3,4,5-triol (cas: 367-93-1) belongs to alcohols. Similar to water, an alcohol can be pictured as having an sp3 hybridized tetrahedral oxygen atom with nonbonding pairs of electrons occupying two of the four sp3 hybrid orbitals. A multistep synthesis may use Grignard-like reactions to form an alcohol with the desired carbon structure, followed by reactions to convert the hydroxyl group of the alcohol to the desired functionality.Synthetic Route of C9H18O5S
Referemce:
Alcohol – Wikipedia,
Alcohols – Chemistry LibreTexts