Monoclonal and polyclonal antibodies compared for radioimmunoassay of somatomedin-C in patients with acromegaly or hypopituitarism.

1987 ◽  
Vol 33 (9) ◽  
pp. 1593-1596 ◽  
Author(s):  
J M Burrin ◽  
J L Paterson ◽  
P S Sharp ◽  
T H Yeo
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Polyclonal Antibody ◽  
Polyclonal Antibodies ◽  
Detection Limits ◽  
Healthy Adults ◽  
Somatomedin C ◽  
Lower Detection

Abstract We used a synthetic recombinant analog of somatomedin-C (Sm-C) to directly compare the performance of polyclonal and monoclonal antibodies for measuring Sm-C in serum of normal persons and in acromegalic or hypopituitary patients. Mean concentrations of Sm-C in healthy adults were 181 (SD 42) micrograms/L as measured with the monoclonal antibody, 194 (SD 61) micrograms/L with the polyclonal antibody. Both antisera gave excellent discrimination between acromegalics and normals. However, the assay with the polyclonal antibody was more sensitive than that with the monoclonal antibody (lower detection limits: 5 vs 100 micrograms/L) and thus better suited for quantifying Sm-C in samples from hypopituitary patients.

Immunofluorescence Assay Using Monoclonal and Polyclonal Antibodies for Detection of Staphylococcal Enterotoxins A in Milk

2019 ◽  
Vol 13 (1) ◽  
pp. 137-145 ◽  
Author(s):  
Tzonka Godjevargova ◽  
Zlatina Becheva ◽  
Yavor Ivanov ◽  
Andrey Tchorbanov
Keyword(s):  
Monoclonal Antibody ◽  
Magnetic Nanoparticles ◽  
Polyclonal Antibody ◽  
Polyclonal Antibodies ◽  
Food Poisoning ◽  
Staphylococcal Enterotoxin ◽  
Staphylococcal Enterotoxin A ◽  
Fluorescence Immunoassay ◽  
Magnetic Separator ◽  
Milk Samples

Objectives: Staphylococcus aureus is a Gram-positive microorganism. S. aureus can grow in various foods and cause food poisoning by secreting enterotoxins. The most common enterotoxins involved in food poisoning are staphylococcal enterotoxin A and staphylococcal enterotoxin B, but Staphylococcal Enterotoxin A (SEA) is predominant. The main types of food contaminated with SEs are meat and meat products, poultry and eggs, milk and dairy products. The aim of this study was to develop a rapid and sensitive fluorescence immunoassay for detection of staphylococcal enterotoxin A in milk. Methods: Monoclonal and polyclonal antibodies for SEA were produced and characterized. Competitive fluorescence immunoassay based on Magnetic Nanoparticles (MNPs) was performed and optimized. MNPs were used as a solid carrier of the antibodies. The first step of the assay was immunoreaction between the immobilized antibody onto MNPs and SEA in milk sample. Then the fluorescein-SEA conjugate was added to the sample. Thus, competitive immunoreaction between MNP-mAb/MNP-pAb with SEA and SEA-FITC was performed. These immuno-complexes were separated by a magnetic separator and the obtained supernatants were analyzed. The fluorescent signal from the excess of conjugated SEA was proportional to the SEA contained in the milk. The assay duration was only 30 min. Results: The fluorescence immunoassays performed with polyclonal antibody had linear ranges from 5 pg/mL to 100 ng/mL SEA in a buffer, and from 50 pg/mL to 50 ng/mL SEA in spiked milk samples. While the same assays performed with monoclonal antibody had linear ranges from 1 pg/mL to 20 ng/mL SEA in buffer, and from 10 pg/mL to 10 ng/mL SEA in spiked milk samples. The detection limits of the developed immunoassays performed in milk were: 48 pg/mL with polyclonal antibody and 9 pg/mL with monoclonal antibody. Conclusion: A rapid and sensitive fluorescence immunoassay based on magnetic nanoparticles with a polyclonal and monoclonal antibody for determination of staphylococcal enterotoxin A in milk was developed.

scholarly journals Circumsporozoite proteins of malaria parasites contain a single immunodominant region with two or more identical epitopes.

1983 ◽  
Vol 157 (6) ◽  
pp. 1947-1957 ◽  
Author(s):  
F Zavala ◽  
A H Cochrane ◽  
E H Nardin ◽  
R S Nussenzweig ◽  
V Nussenzweig
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Specific Binding ◽  
Polyclonal Antibodies ◽  
Binding Assays ◽  
Malaria Parasites ◽  
Immunodominant Region ◽  
Additional Finding ◽  
Single Region ◽  
Competition Binding

We have used panels of monoclonal antibodies to circumsporozoite (CS) proteins of Plasmodium falciparium, P. vivax, and P. knowlesi to determine the number of topographically independent epitopes of these antigens. The results of competition binding assays indicated that single regions of the CS molecules were recognized by the homologous monoclonal antibodies. Competition binding assays were also used to study the specificity of antibodies contained in the sera of humans and monkeys that had developed sterile immunity after immunization with irradiated, intact sporozoites. We found that single monoclonal antibodies inhibited 70-95% of the specific binding of the polyclonal antibodies to crude extracts of sporozoites. It appears, therefore, that CS proteins are among the most immunogenic constituents of sporozoites, and that a single region of these molecules contains most of the immunogenic activity. An additional finding was that the immunodominant region of CS molecules is multivalent with regard to the expression of a single epitope. This was demonstrated by the ability of monomers of CS proteins to bind simultaneously two or more molecules of the same monoclonal antibody.

Abstract 14697: Novel Assays for Quantification of Lipoprotein-Associated (PCSK9-apoB, PCSK9-Lp(a)) Proprotein Covertase Subtilisin/Kexin Type 9 (PCKS9)

Circulation ◽  
2015 ◽  
Vol 132 (suppl_3) ◽  
Author(s):  
Calvin Yeang ◽  
Yun-Seok Choi ◽  
Sang-Rok Lee ◽  
Monica L Bertoia ◽  
Eric B Rimm ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Human Plasma ◽  
Polyclonal Antibodies ◽  
Human Monoclonal Antibody ◽  
Ldl Receptor ◽  
Plasma Lipoproteins ◽  
Health Study ◽  
Clinical Samples ◽  
Ldl Particles

Background: PCSK9 is a major regulator of plasma LDL-C. Monoclonal antibodies to PCSK9 lower LDL-C by 45-65% and Lp(a) by 9-38%. The canonical function of PCSK9 is binding of LDL-receptor (LDLR) via its extracellular EGF-A domain, and subsequently mediating LDLR degradation. However, PCSK9 also weakly associates with plasma lipoproteins, with 20-40% of total plasma PCSK9 found on LDL. However, most LDL particles do not contain PCSK9. Whether PCSK9 also associates with other lipoproteins such as Lp(a) are not well described. Methods: Sensitive and quantitative sandwich-based ELISA assays were developed to measure PCSK9 associated plasma lipoproteins in both mouse and human plasma. For human plasma, commercial rabbit polyclonal antibodies binding to the C-terminal region of PCSK9 (Abgent, ThermoFisher) or REGN727 human monoclonal antibody were bound to microtiter well plates. Plasma was added and monoclonal antibodies MB47 and LPA4, binding to apoB-100 and apo(a) respectively, were used to detect PCSK9-apoB-100 and PCSK9-Lp(a) complexes with a chemiluminescent ELISA. For mouse assays, REGN727 was used as the capture antibody as it detects mouse PCSK9 and monoclonal antibody LF3 was used to detect mouse apoB. Results: PCSK9-apoB and PCSK9-Lp(a) complexes could be detected in both human plasma and in various mouse models expressing apo(a) or Lp(a). The signal to noise ratio was ~20 fold in various clinical samples, including in healthy subjects and in patients with cardiovascular disease. In 536 clinical samples from the Health Professional Follow-Up Study, PCSK9-Lp(a) correlated strongly with Lp(a) (r=0.59, p<0.001, age-adjusted) but not other lipid variables. PCSK9-apoB correlated weakly with PCSK9-Lp(a) (r=0.30, p<0.001, age-adjusted) and LDL-C (r=0.22, p<0.001, age-adjusted). These associations were virtually the same in 526 women in the Nurses’ Health Study. Conclusions: Novel ELISAs were generated to quantitate lipoprotein-associated PCSK9 in transgenic mouse and human plasma, including on apoB and Lp(a). Changes in PCSK9-Lp(a) complexes may provide insights into the Lp(a)-lowering effect of PCSK9 antibodies. Whether these assays will predict CVD outcomes waits to be determined in PCSK9 antibody and epidemiological studies.

scholarly journals Humans surviving cholera develop antibodies against Vibrio cholerae O-specific polysaccharide that inhibit pathogen motility

2020 ◽  
Author(s):  
Richelle C. Charles ◽  
Meagan Kelly ◽  
Jenny M. Tam ◽  
Aklima Akter ◽  
Motaher Hossain ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Vibrio Cholerae ◽  
Polyclonal Antibody ◽  
High Speed ◽  
Polyclonal Antibodies ◽  
Intestinal Lumen ◽  
Dependent Manner ◽  
Specific Antibodies ◽  
Specific Polysaccharide

ABSTRACTThe mechanism of protection against cholera afforded by previous illness or vaccination is currently unknown. We have recently shown that antibodies targeting O-specific polysaccharide (OSP) of Vibrio cholerae correlate highly with protection against cholera. V. cholerae is highly motile and possesses a flagellum sheathed in O-specific polysaccharide (OSP), and motility of V. cholerae correlates with virulence. Using high speed video microscopy, and building upon previous animal-related work, we demonstrate that sera, polyclonal antibody fractions, and OSP-specific monoclonal antibodies recovered from humans surviving cholera block V. cholerae motility at both subagglutinating and agglutinating concentrations. This anti-motility effect is reversed by pre-adsorbing sera and polyclonal antibody fractions with purified OSP; and is associated with OSP-specific but not flagellin-specific monoclonal antibodies. F[ab] fragments of OSP-specific polyclonal antibodies do not inhibit motility, suggesting a requirement for antibody-mediated crosslinking in motility inhibition. We show that OSP-specific antibodies do not directly affect V. cholerae viability, but that OSP-specific monoclonal antibody highly protects against death in the murine cholera model. We used in vivo competitive index studies to demonstrate that OSP-specific antibodies impede colonization and survival of V. cholerae in intestinal tissues, and that this impact is motility-dependent. Our findings suggest that the impedance of motility by antibodies targeting V. cholerae OSP contributes to protection against cholera.IMPORTANCECholera is a severe dehydrating illness of humans caused by Vibrio cholerae. V. cholerae is a highly motile bacterium that has a single flagellum covered in lipopolysaccharide (LPS) displaying O-specific polysaccharide (OSP), and V. cholerae motility correlates with its ability to cause disease. The mechanisms of protection against cholera are not well understood; however, since V. cholerae is a non-invasive intestinal pathogen, it is likely that antibodies that bind the pathogen or its products in the intestinal lumen contribute to protection from infection. Here, we demonstrate that OSP-specific antibodies isolated from humans surviving cholera in Bangladesh inhibit V. cholerae motility and are associated with protection against challenge in a motility-dependent manner.

scholarly journals Two-site immunofluorometric assay of intact salmon calcitonin with improved sensitivity

1997 ◽  
Vol 43 (1) ◽  
pp. 71-75 ◽  
Author(s):  
Haiqin Rong ◽  
Leonard J Deftos ◽  
Hong Ji ◽  
Elisabet Bucht
Keyword(s):  
Monoclonal Antibody ◽  
Detection Limit ◽  
Polyclonal Antibody ◽  
Salmon Calcitonin ◽  
Polyclonal Antibodies ◽  
Pharmacokinetic Studies ◽  
Human Calcitonin ◽  
Plasma Samples ◽  
Serum Concentrations ◽  
Capture Antibody

Abstract We recently developed a two-site immunofluorometric assay (IFMA) of salmon calcitonin (SCT) by DELFIA (dissociation enhancement lanthanide fluoroimmunoassay) technique using the same polyclonal antibodies both for “catching” the antigen and for signaling. In the present study we used a monoclonal antibody to SCT 1–11 as the capture antibody. This antibody was biotinylated before use in streptavidin-coated microtitration plates. The polyclonal antibody labeled with Eu chelate was used as a signaling marker. This combination of antibodies resulted in an assay that was three to four times more sensitive than the previous IFMA, with a detection limit of 0.3 pmol/L serum. Intact SCT 1–32 was detected by the assay (recoveries 94–96%), but not the fragments SCT 1–11 and SCT 10–32 or human calcitonin. Dilutions of plasma samples containing SCT were parallel to the calibration curve of SCT 1–32. Pharmacokinetic studies of SCT, 100 IU administered intramuscularly to 10 men, indicated peak serum concentrations of 32–128 pmol/L within 10–20 min with apparent half-life of 56 ± 18 min (mean ± SD). This new assay will allow study of the pharmacokinetics of new calcitonin preparations that do not require injection.

Evidence for heterogeneity in the 160/165 × 10(3) Mr glycoprotein components of desmosomes

1987 ◽  
Vol 88 (4) ◽  
pp. 513-520
Author(s):  
J.C. Jones ◽  
K.L. Vikstrom ◽  
R.D. Goldman
Keyword(s):  
Monoclonal Antibodies ◽  
Polyclonal Antibody ◽  
Polyclonal Antibodies ◽  
Mouse Skin ◽  
Basal Cells ◽  
Antibody Preparation ◽  
Plaque Component ◽  
Mouse Tissues ◽  
Double Label ◽  
Cryostat Sections

We have prepared both monoclonal and polyclonal antibody preparations directed against the 160/165 × 10(3) Mr glycoproteins (desmogleins) of bovine tongue epithelial desmosomes. The polyclonal antibody preparation recognizes desmosomes in a number of mouse tissues, e.g. mouse skin, heart, bladder and trachea, as determined by immunofluorescence microscopy. Furthermore, the polyclonal antibodies recognize polypeptide(s), present in the high salt, Triton-insoluble residues (‘cytoskeleton preparations’) of mouse skin, heart, bladder and trachea, which comigrate with the 160/165 × 10(3) Mr glycoproteins of bovine tongue epithelial desmosomes as determined by ‘Western’ immunoblotting. Conversely, the monoclonal 160/165 × 10(3) Mr antibody preparation recognizes desmosomes of stratified squamous epithelial tissues but not desmosomes in other tissue types. Moreover, whereas the monoclonal antibodies recognize 160/165 × 10(3) Mr polypeptides in mouse skin cell cytoskeletons they show no immunoreactivity with the cytoskeleton preparations of mouse bladder, trachea and heart following immunoblotting. These results suggest therefore that although there are conserved epitopes of the 160/165 × 10(3) Mr glycoproteins there are also epitopes of these molecules which vary from tissue to tissue. Double label immunofluorescence observations of cryostat sections of mouse skin using the monoclonal antibodies and antibodies directed against desmoplakin, a plaque component of desmosomes, reveal that the monoclonal antibodies do not recognize certain desmosomes in basal cells which are recognized by desmoplakin antibodies. Indeed, double label observations of cryostat sections of mouse skin using the monoclonal antibodies and human autoantibodies which react with hemidesmosomal components suggest that the monoclonal antibodies stain desmosomes located along the apical surfaces of basal cells but fail to recognize desmosomes along the lateral surfaces of these same cells.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals A comparative study of the use of monoclonal antibodies using three different immunohistochemical methods: an evaluation of monoclonal and polyclonal antibodies against human prostatic acid phosphatase.

1982 ◽  
Vol 30 (3) ◽  
pp. 253-260 ◽  
Author(s):  
W Y Naritoku ◽  
C R Taylor
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Acid Phosphatase ◽  
Polyclonal Antibodies ◽  
Prostatic Acid Phosphatase ◽  
Mouse Serum ◽  
Paraffin Sections ◽  
Layer System ◽  
Significant Difference ◽  
Immunohistochemical Methods

The use of immunohistochemical methods has been advocated for the detection and localization of prostatic acid phosphatase in paraffin sections of human prostate. This article explores the possible advantages of utilizing monoclonal antibodies in this method. Monoclonal antibodies, specific for human prostatic acid phosphatase, were integrated into three different immunohistochemical procedures. In the first method, a three-layer peroxidase-antiperoxidase (PAP) system was employed; the monoclonal antibody was followed by rabbit bridge antibody directed against mouse immunoglobulin and mouse PAP complex. The second method was a three-layer system utilizing biotin-labeled horse anti-mouse antibody as "bridge" antiserum between the primary monoclonal antibody and an avidin-biotin-horseradish peroxidase complex. The third method was a four-layer system; the monoclonal antibody was followed by rabbit anti-mouse serum, swine anti-rabbit immunoglobulin as the bridge antibody and rabbit PAP complex. It was found that some, but not all, monoclonal antibodies can be used for the detection of prostatic acid phosphatase in paraffin sections. The four-layer PAP method was found to be the most sensitive method of the three systems tested; however, the avidin-biotin method required the least amount of time. No significant difference in the quality of staining was observed between monoclonal antibodies and carefully absorbed conventional antiserum.

Production of Monoclonal Antibodies Against Enamelin and Against Amelogenin Proteins of Developing Enamel Matrix

1987 ◽  
Vol 1 (2) ◽  
pp. 282-288 ◽  
Author(s):  
D. Deutsch ◽  
A. Palmon ◽  
J. Catalano-Sherman ◽  
R. Laskov
Keyword(s):  
Monoclonal Antibody ◽  
Extracellular Matrix ◽  
Monoclonal Antibodies ◽  
Structural Organization ◽  
Polyclonal Antibodies ◽  
Protein Band ◽  
Enamel Matrix ◽  
Complex Process ◽  
The One ◽  
Successful Production

The extracellular matrix of developing enamel contains two major classes of proteins, the hydrophobic proline-rich amelogenins and the acidic serine-, glycine-, and aspartic-rich enamelins. These proteins have been postulated as playing a major role in the mineralization and structural organization of developing enamel. To identify and further characterize these different proteins and their possible role in this complex process of biological mineralization, we have in recent years been concerned with the production of specific probes for these proteins. Previously, we have reported on the successful production of specific polyclonal antibodies against enamelin proteins, which did not cross-react with amelogenins, and against amelogenin proteins, which did not cross-react with enamelins (Deutsch et al., 1986, 1987). We now report the production of monoclonal antibodies against a major bovine amelogenin protein (28 kDa) and against a major bovine enamelin protein (66 kDa). One monoclonal antibody against amelogenin and one against enamelin are described. The results showed that the monoclonal antibody against the amelogenin protein reacted strongly with the 28-kDa amelogenin protein band but did not cross-react with enamelins, and the one against the enamelin protein reacted with the 66-kDa enamelin protein but did not cross-react with amelogenins. These monoclonal antibodies provide a specific and powerful tool to distinguish between and further characterize these different classes of proteins, and to improve our understanding of the process of enamel formation.

Competitive Direct Enzyme-Linked Immunosorbent Screening Assay for the Detection of Sulfamethazine Contamination of Animal Feeds

1991 ◽  
Vol 74 (5) ◽  
pp. 784-789 ◽  
Author(s):  
Dixon E Holland Deborah ◽  
Stanley E Katz
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Polyclonal Antibodies ◽  
Screening Method ◽  
Screening Assay ◽  
Animal Feeds ◽  
Feed Samples

Abstract A sensitive screening method has been developed for detecting sulfamethazine (SMZ) contamination of feeds by using either polyclonal or monoclonal antibodies and a direct competitive enzyme-linked immunosorbent screening assay (ELISA). Feed samples of 25.0 g are extracted with 0.5N HCI and centrifuged. The extract is adjusted to pH 7.0 with 3.0N NaOH and recentrifuged. This pH-adjusted extract is used in the EUSA. Levels as low as 0.004 μg SMZ/g feed were detected In supplemented extracts by polyclonal antibodies; levels of 0.4 μg SMZ/g feed were detected by a monoclonal antibody.

Intermediate Filament Staining in the Cytologic and Histologic Diagnosis of Canine Skin and Soft Tissue Tumors

1988 ◽  
Vol 25 (5) ◽  
pp. 343-349 ◽  
Author(s):  
C. B. Andreasen ◽  
E. A. Mahaffey ◽  
J. R. Duncan
Keyword(s):  
Monoclonal Antibody ◽  
Soft Tissue ◽  
Polyclonal Antibody ◽  
Polyclonal Antibodies ◽  
Soft Tissue Tumors ◽  
Histologic Diagnosis ◽  
Background Staining ◽  
Canine Tumors ◽  
Formalin Fixed ◽  
Avidin Biotin Complex

Formalin-fixed histologic and acetone-fixed cytologic preparations from 87 surgically removed subcutaneous and soft tissue canine tumors were examined for immunoreactivity to cytokeratin. desmin. and vimentin. The avidin-biotin complex (ABC) technique demonstrated immunoreactivity in both preparations. but the intensity and specificity of the reactions were dependent on the primary' antibody. Polyclonal antibodies to cytokeratin were more consistent in immunoreactivity than were polyclonal desmin or vimentin antibodies. The monoclonal antibody proved more satisfactory for demonstrating vimentin than the polyclonal antibody. Greater dilutions of primary antibodies may be used on cytologic preparations than on histologic sections. Evaluation of cytologic preparations may be inconclusive due to background staining, scant cellularity, or poor cytoplasmic preservation.

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