The method of monoclonal antibody-based immunoassay has a great importance in the study of quality control of traditional Chinese medicine (TCM) and detection of trace components in vivo animals. Synthesis of small molecule artificial antigen is the prerequisite for the establishment of this method. In present study, catalpol-BSA was synthesized by sodium periodate oxidation method. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry ( MALDI-TOF-MS) and molecular exclusion chromatography showed that catalpol was successfully conjugated with BSA. The mice could specifically produce anti-catalpol antibodies with titer up to 1:8000. The artificial antigen of catalpol was successfully synthesized.
Animals ; Antibodies ; immunology ; Antigens ; chemistry ; immunology ; Immunoassay ; Iridoid Glucosides ; chemistry ; immunology ; Male ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred BALB C ; Serum Albumin, Bovine ; chemistry ; immunology

OBJECTIVETo explore the effect of Peitu Shengjin Recipe (PSR) on nutritional states and immune functions of stable phase chronic obstructive pulmonary disease (COPD) patients.

METHODSTotally 62 stable phase COPD patients were randomly assigned to the treatment group (30 cases) and the control group (32 cases). All patients inhaled Seretide (50/500 µg), twice per day. Besides, patients in the treatment group additionally received PSR, one dose per day. After three months of treatment, the COPD assessment test (CAT) score, the index of nutritional states [including body mass index (BMI) , thickness of skin fold (TSF), mid-arm muscle circumference (MAMC), serum albumin, serum prealbumin], and immune functions (including IgA, IgM, and IgG) were compared between the two groups.

RESULTSBy the end of the treatment, the CAT score decreased more obviously in the treatment group than in the control group (P < 0.05). The improvement of BMI, TSF, MAMC, serum albumin, and serum prealbumin was better in the treatment group than in the control group (P < 0.05). IgM and IgG also increased more in the treatment group (P < 0.05). There was no statistical difference in IgA between the two groups (P > 0.05).

CONCLUSIONSAdditionally use of PSR could improve nutritional states and immune functions of stable phase COPD patients to some extent. Meanwhile, it also could improve their health related quality of life.

Body Mass Index ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Nutritional Status ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; immunology ; Quality of Life ; Serum Albumin

Oxidation method with sodium iodide was used to synthesize immunogenic antigen (PF-BSA) and coating antigen (PF-OVA) of paeoniflorin. UV spectroscopy showed that paeoniflorin was successfully conjugated with BSA and OVA. After immunized by PF-BSA, the mice can produce anti-paeoniflorin antibodies specifically. The ELISA test results showed the high titer (1:12 800) and specificity (IC50 = 0.791 mg x L(-1)) of the antiserum from mice injected with PF-BSA. Also, the antiserum showed low cross activities against nine traditional Chinese medicine (TCM) of small molecules. These artificial antigens were successfully synthesized and the anti-paeoniflorin antibody well prepared, which provides the experimental basis for the further study of ELISA and its kit.
Animals ; Antibodies ; analysis ; Antigens ; chemistry ; immunology ; Drugs, Chinese Herbal ; chemistry ; Enzyme-Linked Immunosorbent Assay ; Glucosides ; chemistry ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Monoterpenes ; chemistry ; immunology ; Serum Albumin, Bovine ; chemistry ; immunology

OBJECTIVESynthesis and identification of complete antigen of rutin, the traditional Chinese medicine active ingredient, and develop rapid detection of rutin using enzyme-linked immunoassay method (ELISA). Immunogenicity of the complete antigen was also studied.

METHODPrepare the complete antigen by sodium periodate solution and identified by UV scanning and SDS-PAGE test. Male New Zealand white rabbits were immunized by the antigen to obtain the antiserum.

RESULTThe results of UV analysis showed that the coupling ratio of complete antigen is 13: 1. SDS-PAGE display of the artificial antigen was delayed compared with bovine serum protein. The titer of rutin antibody is 1:4 000. The sensitivity of IC50 was 5.37 mg x L(-1), the lowest detection limit was 1 mg x L(-1), the average recovery was 102%, the intra and interspecific RSD were less than 10%, cross-reactivity rate of antibodies and other analogs were less than 1%.

CONCLUSIONRutin complete antigen was synthesized successfully, and the rapid detection of rutin by ELISA method was successfully established.

Animals ; Antibody Specificity ; immunology ; Antigens ; immunology ; Cattle ; Cross Reactions ; immunology ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Immune Sera ; immunology ; Immunization ; Male ; Periodic Acid ; chemistry ; Rabbits ; Rutin ; chemical synthesis ; immunology ; Serum Albumin, Bovine ; immunology ; Solutions ; chemistry

There are some small molecules with potential allergenicity in traditional Chinese medicine injections. They are lack of immunogenicity due to their small molecular weight, but they can lead to allergic reactions when they were coupled with appropriate vectors. Therefore, how to couple small molecule semi-antigens with vectors to prepare complete antigens with immunogenicity and reactogenicity is the key for screening small molecular allergenic substances out of traditional Chinese medicine injections. In terms of semi-antigen characteristics of traditional Chinese medicine injections, vector selection and application, coupling method and complete antigen purification and identification, the author introduces the latest research situations of artificial antigen and antibody preparation technology, the advance in experimental studies on screening of allergenic substances in traditional Chinese medicine injections, as well as the application prospect of immuno-chip technology in studies on allergenic substances in traditional Chinese medicine injections, with the aim of providing new experimental thoughts and methods for safety control of traditional Chinese medicine injections.
Allergens ; administration & dosage ; chemistry ; immunology ; Antigens ; administration & dosage ; chemistry ; immunology ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Humans ; Hypersensitivity ; immunology ; Injections ; Medicine, Chinese Traditional ; methods ; trends ; Serum Albumin ; administration & dosage ; chemistry ; immunology

A monoclonal antibody (mAb) against zearalenone (ZEN) was produced using ZEN-carboxymethoxylamine and -BSA conjugates. Antibody produced by one clone showing a very high binding ability was selected and found to have a higher affinity for ZEN compared to a commerciall ZEN antibody. We developed two direct competitive ELISA systems using the selected antibody (ZEN-coated and anti-ZEN antibody-coated ELISA). Quantitative ranges for the anti-ZEN antibody-coated ELISA and ZEN-coated ELISA were from 25 to 750 ppb and from 12.5 to 100 ppb, respectively. The detection limit of both methods as measured with standard solutions was 10 ppb. The intra-plate and inter-well variation of both ELISAs were less than 10%. The IC50 values for alpha-zearalenol, beta-zearalenol, alpha-zearalanol, and beta-zearalanol compared to ZEN were 108.1, 119.3, 114.1, and 130.3% for the ZEN-coated ELISA. These values were 100.7, 120.7, 121.6, and 151.6% for the anti-ZEN antibody-coated ELISA. According to the anti-ZEN antibody-coated ELISA, the average recovery rates of ZEN from spiked animal feed containing 150 to 600 ng/mL of ZEN ranged from 106.07 to 123.00% with 0.93 to 2.28% coefficients of variation. Our results demonstrate that the mAb developed in this study could be used to simultaneously screen for ZEN and its metabolites in feed.
Aminooxyacetic Acid/*chemistry ; Animals ; Antibodies, Monoclonal/*immunology ; Enzyme-Linked Immunosorbent Assay/*methods ; Female ; Inhibitory Concentration 50 ; Mice ; Mice, Inbred BALB C ; Reproducibility of Results ; Serum Albumin, Bovine/chemistry ; Zearalenone/*immunology

This is the first reported detection of serum IgE antibody to piperacillin-human serum albumin (HSA) conjugate in a patient presenting with anaphylaxis that developed after occupational exposure. A 24-yr-old nurse, who had worked at a University Hospital for 2 yr, experienced chest tightness, dizziness, generalized urticaria, abdominal pain, and diarrhea 10 min after administering a piperacillin injection. She had previously suffered from atopic dermatitis. A skin prick test for common inhalant allergens was entirely negative; in contrast, her serum total IgE was elevated (283 IU/mL). A high level of piperacillin-specific serum IgE was detected by ELISA using piperacillin-HSA conjugate. Significant inhibition upon addition of both free piperacillin and piperacillin-HSA conjugate was detected by inhibition ELISA. These data suggest that piperacillin exposure in the workplace can induce occupational anaphylaxis and urticaria mediated by an interaction of IgE with the hapten of piperacillin.
Anaphylaxis/*chemically induced/immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Hospitals, University ; Humans ; Immunoglobulin E/*blood/immunology ; Intensive Care Units ; Occupational Diseases/*chemically induced/immunology ; *Occupational Exposure ; Piperacillin/*immunology ; Serum Albumin/*immunology ; Urticaria/immunology ; Young Adult

OBJECTIVETo identify and explore the prognostic risk factors of the hemophagocytic syndrome (HPS).

METHODSA retrospective study was conducted on 50 childhood patients with HPS who were admitted to our hospital between 2007 and 2011. All their medical records were reviewed and analyzed. For each patient, demographic, laboratory data and outcome information were collected. The patients were divided into deceased or survived groups based on the follow-up results. Comparative analysis of the data was done by using independent-samples test and logistic multiple and univariate regression.

RESULTSAmong the 50 HPS patients, 30 were male and 20 female, age ranged from 3 months to 10 years. Reduction of serum albumin, cholinesterase and natural killer (NK) cells was found in the forty-six patients. The laboratory features showed an elevation of serum ferritin with hypofibrinogenemia and hypertriglyceridemia in most of the patients. Forty of patients had hemophagocyte in bone marrow at diagnosis of HPS. The positive serum EBV-IgM was found in thirty-five patients.During the observation period, 25 of 37 patients (67.6%) died, while 13 of whom died within a month after hospitalization. The deceased patients were more likely to have lower albumin, cholinesterase, NK cells level and more prolonged active partial thromboplastin time than the survived patients (P < 0.05). Multivariate logistic regression analysis revealed that duration of illness > 1 month, albumin level < 25 g/L, cholinesterase level < 2000 U/L, NK cell level 0-3% and positive EBV-IgM were related with the prognosis significantly (P < 0.05 for all comparisons).

CONCLUSIONThis study revealed that duration of illness > 1 month, decreases in albumin, NK cell and cholinesterase, and positive EBV-IgM were the risk factors related to mortality in children.

Child ; Child, Preschool ; Cholinesterases ; blood ; Female ; Herpesvirus 4, Human ; immunology ; Humans ; Immunoglobulin M ; blood ; Infant ; Killer Cells, Natural ; immunology ; Lymphohistiocytosis, Hemophagocytic ; diagnosis ; mortality ; pathology ; Male ; Prognosis ; Retrospective Studies ; Risk Factors ; Serum Albumin ; analysis ; Syndrome

To synthesize salbutamol immunogen and develop an enzyme immunoassay (ELISA), a new salbutamol immunogen was synthesized using 4-aminobenzoic acid as a linker to connect hapten with carrier protein. An enzyme immunoassay based on the antibody prepared was developed and applied to detect salbutamol residue spiked in swine liver. An unusual coating antigen, clenbuterol-ovalbumin (OVA) conjugate instead of salbutamol-OVA conjugate, was used in the immunoassay and the results were discussed based on the structures of related compounds. The antibodies showed high sensitivity in the heterologous assay when using clenbuterol-OVA as a coating antigen, with an IC50 value of 8.97 ng mL(-1) toward salbutamol. The antibodies prepared showed high cross-reactivity with clenbuterol (107%) and were promising for the simultaneous determination of salbutamol and clenbuterol residues in food and food products. Recovery rates from the salbutamol-spiked swine liver samples were in the range of 70%-99%, while the intra-assay and inter-assay coefficients of variation were <13.3% and <14.3%, respectively. In summary, the antibodies of salbutamol have been successfully prepared. Sensitive and stable analysis for the detection of salbutamol residues in swine liver was obtained based on the competitive ELISA methods developed in this study.
4-Aminobenzoic Acid ; chemistry ; Adrenergic beta-Agonists ; analysis ; immunology ; Albuterol ; analysis ; immunology ; Animals ; Antibodies ; immunology ; Antibody Specificity ; Clenbuterol ; analysis ; immunology ; Drug Residues ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Food Contamination ; Haptens ; immunology ; Immunization ; Liver ; chemistry ; Male ; Ovalbumin ; chemistry ; immunology ; Rabbits ; Serum Albumin, Bovine ; chemistry ; immunology ; Swine

It is generally believed that liposomes modified with polyethylene glycol (PEG) have no or lower immunogenicity. However, based on many recent literatures, when the PEGylated liposomes were repeatedly applied to the same animal, the immune responses occurred. The first injection of PEGylated liposomes resulted in a reduction in the circulation time and an increase in hepatic and splenic accumulation of the second dose of PEGylated liposomes in a time-interval, which was called "accelerated blood clearance (ABC)" phenomenon. Such immunogenicity of PEGylated liposomes presents a barrier in the research of liposomal formulations and their use in the clinics. This review focused on the definition, the method of verification, the development of the reason for ABC phenomenon, influencing factors of ABC phenomenon, and discussed if other PEGylated nanocarriers also induce ABC phenomenon.
Animals ; Antibiotics, Antineoplastic ; administration & dosage ; pharmacokinetics ; Doxorubicin ; administration & dosage ; pharmacokinetics ; Drug Carriers ; Immunoglobulin M ; biosynthesis ; blood ; Liposomes ; administration & dosage ; blood ; pharmacokinetics ; Liver ; metabolism ; Metabolic Clearance Rate ; Particle Size ; Polyethylene Glycols ; administration & dosage ; metabolism ; pharmacokinetics ; Serum Albumin, Bovine ; pharmacokinetics ; Spleen ; immunology ; metabolism