Two desired epitopes of cTnI benefit for preparation of standardized monoclonal antibodies.

Acute myocardial infarction (AMI) is one of the most severe cardiovascular diseases in humans, often resulting in unexpected death. Early detection is critical for patient survival. Sandwich ELISA is a common method for the detection of AMI.

Plasmonic ELISA for Sensitive Detection of Disease Biomarkers with a Smart Phone-Based Reader.

Serum myoglobin is one of the earliest markers for the diagnosis of acute myocardial infarction. It is, therefore, critical to develop a point-of-care testing technology for myoglobin detection. In this work, we reported a sensitive plasmonic immunoassay-based on enzyme-mediated localized surface plasmon resonance change of gold nanorods for the point-of-care testing detection of myoglobin.

Construction of a human monoclonal antibody against bFGF for suppression of NSCLC.

Compelling evidence implicates that overexpression of basic fibroblast growth factor (bFGF) and fibroblast growth factor receptor 1 (FGFR1) in non-small cell lung cancer (NSCLC) drives tumor progression, can serve as prognostic biomarkers or therapeutic targets for NSCLC patients. But at present, we still lack of effective drugs for bFGF. The preparation of monoclonal antibodies against bFGF or to understand its mechanism of action is urgently need.

[Effect of basic fibroblast growth factor antibody combined with irinotecan on proliferation and apoptosis of small cell lung cancer H223 cells in vitro].

Objective: To study the synergistic inhibitory effects of basic fibroblast growth factor (bFGF) monoclonal antibody (bFGF mAb) and irinotecan on the proliferation of small cell lung cancer H223 cells.

Methods: CCK-8 assay and flow cytometry were used to assess the effects of bFGF mAb combined with irinotecan on the proliferation and apoptosis of H223 cells, respectively. Western blotting was performed to analyze the effect of bFGF-mAb combined with irinotecan on AKT and ERK1/2 phosphorylation in the cells.

BFGF neutralization stimulates VEGF secretion in melanoma B16 cells.

Fusarium root rot is a major cryptogamic disease in olive trees caused by the soil-borne fungus Fusarium solani. Controlling this disease requires the extensive use of chemicals. However, using BCAs such as some Trichoderma strains may be an opportune alternative to fungicides in protecting olive plantations.

(125)I-labeled anti-bFGF monoclonal antibody inhibits growth of hepatocellular carcinoma.

Aim: To investigate the inhibitory efficacy of (125)I-labeled anti-basic fibroblast growth factor (bFGF) monoclonal antibody (mAb) in hepatocellular carcinoma (HCC).

Methods: bFGF mAb was prepared by using the 1G9B9 hybridoma cell line with hybridization technology and extracted from ascites fluid through a Protein G Sepharose affinity column. After labeling with (125)I through the chloramine-T method, bFGF mAb was further purified by a Sephadex G-25 column.

Development of SPR biosensor for simultaneous detection of multiplex respiratory viruses.

A surface plasmon resonance (SPR)-based biosensor was developed for specific detection of nine common respiratory virus, including influenza A and influenza B, H1N1, respiratory syncytial virus (RSV), parainfluenza virus 1-3 (PIV1, 2, 3), adenovirus, and severe acute respiratory syndrome coronavirus (SARS). The SPR biosensor was developed by immobilizing nine respiratory virus-specific oligonucleotides in an SPR chip. To increase the biosensor sensitivity, biotin was used to label the PCR primer and further amplify the signal by introducing streptavidin after hybridization.

[Preparation and application of monoclonal antibody against human myoglobin].

Objective: To prepare the monoclonal antibody (mAb) against human myoglobin (MYO) of high titer and specificity and develop double-antibody sandwich ELISA for detecting MYO in human serum samples.

Methods: The BALB/c mice were immunized with natural human MYO, and the hybridoma cell lines secreting anti-MYO mAb were established using cell fusion and hybridoma screening techniques. The characteristics of the mAb were identified after affinity purification from ascites.

A novel method to detect Listeria monocytogenes via superparamagnetic lateral flow immunoassay.

A novel strip test system combining immunomagnetic separation with lateral flow immunoassay (LFIA) was established for the accurate detection of Listeria monocytogenes. In this system, a pair of matched monoclonal antibodies was used to construct a sandwich immunoassay, in which superparamagnetic particles were coupled with one of the antibodies as a labeled antibody to capture the target bacteria, while the other antibody was immobilized on the detection zone. After a 20-min reaction, the strips were analyzed by a novel instrument which could detect the magnetic signal of the immunocomplex in a magnetic field.

[Rapid detection of Listeria monocytogenes by immunomagnetic separation combined with selective medium].

Listeria monocytogenes is a pathogenic bacterium, therefore, it is essential for food safety monitoring to establish a rapid and specific detecting method. In this study, immunomagnetic beads and selective medium were combined to detect Listeria monocytogenes at different concentrations (10(1)-10(5) CFU/mL). Other three types of Listeria spp.

[Preparation of anti-zearalenone monoclonal antibody and preliminary establishment of colloidal gold immunochromatographic assay for zearalenone].

Objective: To prepare the monoclonal antibody against zearalenone (ZEN) toxin and preliminarily establish the colloidal gold immunochromatographic detection method for ZEN.

Methods: The artificial antigen ZEN-BSA and ZEN-OVA were prepared by active ester method. Mice were immunized with ZEN-OVA and monoclonal antibodies against ZEN were prepared by regular cell fusion and subcloning approach.

A novel fluorescence-quenching immunochromatographic sensor for detection of the heavy metal chromium.

A novel fluorescence quenching immunochromatographic sensor (ICS) was developed for detecting chromium (Cr(3+)) within 15 min utilizing the fluorescence quenching function of gold nanoparticles (Au-NPs). The sensor performed with a positive readout. When the low concentrations of Cr(3+) samples were applied, detection signals of the test line (T line) were quenched, whereas when higher concentration Cr(3+) samples (1.

Colloidal gold nanoparticle probe-based immunochromatographic assay for the rapid detection of chromium ions in water and serum samples.

An immunochromatographic assay (ICA) using gold nanoparticles coated with monoclonal antibody (McAb) for the detection of chromium ions (Cr) in water and serum samples was developed, optimized and validated. Gold nanoparticles coated with affinity-purified monoclonal antibodies against isothiocyanobenzyl-EDTA (iEDTA)-chelated Cr(3+) were used as the detecting reagent in this completive immunoassay-based one-step test strip. The ICA was investigated to measure chromium speciation (Cr(3+) and Cr(6+) ions) in water samples.

[Mass spectrometry identification and immune cross-reactivity of a minor shrimp allergen-sarcoplasmic calcium binding protein from Litopenaeus vannamei].

Aim: To identify sarcoplasmic calcium-binding protein (SCP) as a minor shrimp allergen by mass spectrometry, and to analyze the immune cross-reactivity among crustacean SCPs.

Methods: The M(r); 21 000 allergen from Litopenaeus vannamei was identified by MALDI-TOF/TOF-MS. BLAST and ClustalW were used to compare amino acid sequence identity of the allergen among crustaceans.

Development of a lateral flow immunoassay (LFA) strip for the rapid detection of 1-aminohydantoin in meat samples.

Unlabelled: Due to the potential toxic effects of the nitrofuran family of antibiotics, their use in animals in the food industry has raised health concerns. This study was aimed to develop a lateral flow assay (LFA) based on competitive format for the detection of 1-aminohydantoin (AHD) in meat samples. The assay could be completed in 1 min and detected AHDs derivates (CPAHD) at 3 ng/mL, equivalent to 1.

[Preparation and application of monoclonal antibodies against Heavy Metal Chromium].

Aim: To produce the monoclonal antibodies (mAb) against Heavy Metal Chromium, and develop a competitive inhibition enzyme-linked immunosorbent assay (ciELISA) for the detection of Chromium.

Methods: Chromium ions was chelated with bifunctional chelating agent Isothio-cyanobenzyl- EDTA firstly and then conjugated with bovin serum alburmin(BSA) and ovalbmin(OVA) respectively to complete antigens . Four female BALB/c mice were immunized with Cr-iEDTA-BSA, the Hybridoma lines secreting monoclonal antibodies against Chromium ions were established by the hybridoma technology.

Monoclonal antibodies targeting basic fibroblast growth factor inhibit the growth of B16 melanoma in vivo and in vitro.

Up-regulated basic fibroblast growth factor (bFGF or FGF-2) plays an important role in the development and metastasis of melanoma; therefore, neutralizing antibodies to bFGF may suppress melanoma growth. In this study, we have developed three monoclonal antibodies against bFGF (anti-bFGF mAbs), which display remarkable anti-tumor and anti-angiogenic effects in vitro and in vivo. Anti-bFGF mAbs significantly inhibit the proliferation and induce apoptosis of B16 cells, and show inhibitory effects on the migration of B16F10 cells and the tube formation of human umbilical vein endothelial cells (HUVECs) in vitro.

[A Paralytic shellfish poisoning GTX2,3 mimic peptide isolated from phage display: characterization and potential applications].

Aim: To screen peptide mimics of PSP (Paralytic shellfish poisoning) GTX2,3 from a random 12-mer phage display peptide library and to identify and characterize the specificity and accuracy of the peptide mimotope of GTX2,3.

Methods: The monoclonal antibody against GTX2,3 (mAb E(9);F(10);) was used as a target to screen the 12-mer phage display peptide library and the specificity of phage clones were identified by sandwich ELISA and blocking assay. The peptide sequences of positive phage clones were determined and analyzed by DNA sequencing.

[Identification, isolation, purification of allergens in shrimp and allergic analysis for the main allergens].

Aim: To identify the allergens in shrimp, and to isolate, purify and analyze the main allergen components.

Methods: The total shrimp proteins were extracted by PBS, the allergens were identified with 11 shrimp allergic patients' serum IgE by Western blot. Three main shrimp allergens 21,000, 36,000 and 80,000 were purified by ammonium sulfate precipitation, Sephadex G-50 chromatography and DEAE-exchange chromatography.

Colloidal gold probe-based immunochromatographic assay for the rapid detection of lead ions in water samples.

One-step immunochromatographic assay (ICA) has been developed using colloidal gold-labeled monoclonal antibody probe for the rapid detection of lead ions in water samples. The ICA was based on the theory of competitive reactivity, and the results can be easily judged based on the presence or absence of a red colored test line with visual detection. Under optimal conditions, this method shows high detecting sensitivity with a LOD (limit of detection) of 50 ng/ml.

Selection and characterization of a human neutralizing antibody to human fibroblast growth factor-2.

Compelling evidences suggest that fibroblast growth factor-2 (FGF-2) plays important roles in tumor growth, angiogenesis and metastasis. Molecules blocking the FGF-2 signaling have been proposed as anticancer agents. Through screening of a human scFv phage display library, we have isolated several human single-chain Fv fragments (scFvs) that bind to human FGF-2.

A competitive indirect enzyme-linked immunoassay for lead ion measurement using mAbs against the lead-DTPA complex.

Immunoassays for quantitative measurement of environmental heavy metals offer several advantages over other traditional methods. To develop an immunoassay for lead, Balb/c mice were immunized with a lead-chelate-protein conjugate to allow maximum exposure of the metal to the immune system. Three stable hybridoma cell lines were obtained through spleen cells fusion with Sp2/0 cells.

A sensitive immunosorbent bio-barcode assay combining PCR with icELISA for detection of gonyautoxin 2/3.

In the current study, we developed a nanosphere bio-barcode technology to detect trace gonyautoxin 2/3 (GTX 2/3). GTX 2/3-glucose oxidase (GOX) conjugates were first prepared as the coating antigen in a periodate reaction. Subsequently, gold nanoparticles (NP) dual-labeled with anti-GTX 2/3 monoclonal antibodies (Mab) and DNA oligonucleotides were synthesized via a one-step preparation method.

[Cloning of the variable region genes from hybridoma against bFGF and expression of single chain antibody fragments in E.coli HB2151].

Aim: To construct and express the single chain antibody (scFv) in E.coli HB2151 by cloning the variable region genes from hybridoma against bFGF.

Methods: Total RNA was extracted from hybridoma cell line B2F3 secreting mAbs against bFGF and the cDNA was amplified by retropolymerase chain reaction (RT-PCR).

[Expression of human anti-HBsAg single chain Fab gene in Pichia pastoris].

Aim: To investigate the relation of the affinity activity to the structure of recombinant Fab.

Methods: The human anti-HBsAg single chain Fab gene was obtained by overlaping PCR from the H and L chains of Fab, and was introduced into the expression system of Pichia pastoris. The expression vector of single chain Fab was transformed into GS115 cells by the method of LiCl transformation.