Immunoaffinity chromatography of bovine FSH using monoclonal antibodies

ABSTRACT Bovine FSH (bFSH) was used to immunize BALB/c mice. Spleen cells were fused to the SP 2/0 cell line to produce hybridomas that secreted monoclonal antibodies to bFSH. One of these antibodies (USDA-bFSH-MC28) was extensively characterized and found to be a gamma 1 with kappa light chains, having extremely low cross-reactivity with other bovine pituitary hormones and with ovine and porcine FSH. The dissociation constant as measured by Scatchard analysis was 4·3 nmol/l, and proved to be in a very useful range for affinity chromatography. In an essentially one-step immunoaffinity chromatography procedure, bFSH was easily isolated in a single chromatographic step from crude anterior pituitary homogenate with better yield and with the same purity as classical chromatographic techniques. J. Endocr. (1987) 115, 283–288

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Immunological studies on staphylococcal enterotoxin D: production of murine monoclonal antibodies and immunopurification

Canadian Journal of Microbiology ◽

10.1139/m91-099 ◽

1991 ◽

Vol 37 (8) ◽

pp. 586-589

Author(s):

Kunihiro Shinagawa ◽

Katsuhiko Omoe ◽

Naonori Matsusaka ◽

Shunji Sugii

Keyword(s):

Staphylococcus Aureus ◽

Monoclonal Antibodies ◽

Key Words ◽

Immunoaffinity Chromatography ◽

Staphylococcal Enterotoxin ◽

Mouse Spleen ◽

Spleen Cells ◽

Myeloma Cells ◽

Murine Monoclonal Antibodies

Eight murine monoclonal antibodies (MAbs) against staphylococcal enterotoxin D (SED) were obtained by fusion of myeloma cells with mouse spleen cells immunized with SED only or a combination of SED and either enterotoxin A (SEA) or enterotoxin E (SEE). When only SED was used as an immunogen, six MAbs were specific for SED only, whereas one MAb was reactive with both SED and SEE when both SEs were used as immunogens. One MAb reacted with SEA, SED, and SEE when both SEA and SED were used as immunogens. A MAb with the highest reactivity to SED was used to prepare an immunosorbent for purification of SED by immunoaffinity chromatography. Approximately 70% of the partially purified SED was recovered in the eluate. The purified SED was electrophoretically and antigenically pure. Immunoaffinity chromatography proved useful in the purification of SED in terms of ease of purification, percent enterotoxin, and enterotoxin purity. Key words: enterotoxin D, monoclonal antibodies, Staphylococcus aureus.

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The production and characterization of monoclonal antibodies to the human thyroid microsome

Journal of Endocrinology ◽

10.1677/joe.0.1050047 ◽

1985 ◽

Vol 105 (1) ◽

pp. 47-NP ◽

Cited By ~ 9

Author(s):

A. P. Weetman ◽

C. A. Gunn ◽

D. P. Rennie ◽

R. Hall ◽

A. M. McGregor

Keyword(s):

Monoclonal Antibodies ◽

Liver Microsomes ◽

Cross Reactivity ◽

Immunoglobulin M ◽

Human Thyroid ◽

Thyroid Cell ◽

Spleen Cells ◽

Cell Monolayers ◽

Mouse Myeloma

ABSTRACT By suitable immunization of mice and fusion of their spleen cells with a non-secretor mouse myeloma line, monoclonal antibodies have been produced which react with the human thyroid microsomal (M) antigen. These monoclonal antibodies showed no reactivity by enzyme-linked immunoassay with liver microsomes or thyroglobulin and their specificity was confirmed by immunolocalization studies, in which they showed the staining characteristics of human M antibodies. All four monoclonal antibodies tested were immunoglobulin M; three were cytotoxic to thyroid cell monolayers. The lack of cytotoxicity with the fourth monoclonal supports the concept that certain epitopes of the M antigen may be partially or completely absent at the thyroid cell surface. These monoclonal antibodies should permit further characterization of the thyroid M antigen in view of their absence of cross-reactivity with thyroglobulin. J. Endocr. (1985) 105, 47–52

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One-Step Direct Assay for Mature-type Adrenomedullin with Monoclonal Antibodies

Clinical Chemistry ◽

10.1093/clinchem/45.2.244 ◽

1999 ◽

Vol 45 (2) ◽

pp. 244-251 ◽

Cited By ~ 53

Author(s):

Hideki Ohta ◽

Tetsuo Tsuji ◽

Shigeru Asai ◽

Kazuyuki Sasakura ◽

Hiroshi Teraoka ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Healthy Subjects ◽

Plasma Sample ◽

Cross Reactivity ◽

Carboxy Terminus ◽

Large Numbers ◽

Simplified Method ◽

Mature Type ◽

One Step ◽

The Mean

Abstract Adrenomedullin (AM) is a potent hypotensive peptide. Plasma contains mature-type AM (m-AM), which is amidated at the carboxy terminus, and an intermediate, AM-Gly. We developed a one-step two-site IRMA specific for determining human m-AM with monoclonal antibodies. The detection limit was 0.5 pmol/L, and the working range (CV <15%) was 1–300 pmol/L. Dilution of plasma samples showed good linearity. The recovery of added AM was 91–118%. The intra- and interassay imprecision values (CVs) were 4.4–8.2% and 5.5–8.3%, respectively. The assay had no cross-reactivity with AM-Gly or other peptides similar to AM. The mean (± SD) plasma human m-AM concentration of 61 healthy subjects was 1.18 ± 0.65 pmol/L. In conclusion, our IRMA makes it possible to specifically measure m-AM, using a small amount of plasma sample (0.2 mL) by a one-step overnight assay without prior extraction. Our simplified method would be suitable for clinical studies on AM, especially when large numbers of samples must be processed.

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Development of a Colloidal Gold Immunochromatographic Assay for Duck Enteritis Virus Detection Using Monoclonal Antibodies

Pathogens ◽

10.3390/pathogens10030365 ◽

2021 ◽

Vol 10 (3) ◽

pp. 365

Author(s):

Fengli Liu ◽

Yanxin Cao ◽

Maokai Yan ◽

Mengxu Sun ◽

Qingshui Zhang ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Colloidal Gold ◽

Preventive Intervention ◽

Virus Detection ◽

Cross Reactivity ◽

Duck Enteritis Virus ◽

Detection Sensitivity ◽

Immunochromatographic Assay ◽

Efficient Detection ◽

Enteritis Virus

Duck viral enteritis is a highly contagious and fatal disease of commercial waterfowl flocks. The disease occurs sporadically or epizootically in mainland China due to insufficient vaccinations. Early and rapid diagnosis is important for preventive intervention and the control of epizootic events in clinical settings. In this study, we generated two monoclonal antibodies (MAbs) that specifically recognized the duck enteritis virus (DEV) envelope glycoprotein B and tegument protein UL47, respectively. Using these MAbs, a colloidal gold-based immunochromatographic assay (ICA) was developed for the efficient detection of DEV antigens within 15 min. Our results showed that the detection limit of the developed ICA strip was 2.52 × 103 TCID50/mL for the virus infected cell culture suspension with no cross-reactivity with other pathogenic viruses commonly encountered in commercially raised waterfowl. Using samples from experimentally infected ducks, we demonstrated that the ICA detected the virus in cloacal swab samples on day three post-infection, demonstrating an 80% concordance with the PCR. For tissue homogenates from ducks succumbing to infection, the detection sensitivity was 100%. The efficient and specific detection by this ICA test provides a valuable, convenient, easy to use and rapid diagnostic tool for DVE under both laboratory and field conditions.

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Monoclonal antibodies to bovine UDP-galactosyltransferase. Characterization, cross-reactivity, and utilization as structural probes.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(19)57498-1 ◽

1986 ◽

Vol 261 (17) ◽

pp. 7975-7981

Author(s):

J T Ulrich ◽

J R Schenck ◽

H G Rittenhouse ◽

N L Shaper ◽

J H Shaper

Keyword(s):

Monoclonal Antibodies ◽

Cross Reactivity ◽

Structural Probes

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Rapid and simultaneous purification of aflatoxin B1, zearalenone and deoxynivalenol using their monoclonal antibodies and magnetic nanoparticles

Toxicological Research ◽

10.1007/s43188-020-00083-w ◽

2021 ◽

Author(s):

Hyuk-Mi Lee ◽

Hwan-Goo Kang

Keyword(s):

Monoclonal Antibodies ◽

Magnetic Nanoparticles ◽

Aflatoxin B1 ◽

Animal Feed ◽

Immunoaffinity Chromatography ◽

The Novel ◽

Buffer Solution ◽

Purification Method ◽

Recovery Rates ◽

Single Spike

AbstractTo develop a new simple and simultaneous purification method for mycotoxins in feeds and grains, magnetic nanoparticles (MNPs) conjugated with monoclonal antibodies (mAbs) against mycotoxins were used to separate aflatoxin B1 (AFB1), zearalenone (ZEA) and deoxynivalenol (DON). For a single spike of each mycotoxin into the buffer solution (16% MeOH in PBS), mean recoveries were 93.1–95.0% for AFB1 (5–20 ng/mL spiked), 87.2–96.0% for ZEA (125–500 ng/mL spiked) and 75.2–96.9% for DON (250–1,000 ng/mL spiked) by HPLC and ELISA. Recoveries of AFB1 (20 ng/mL) and ZEA (500 ng/mL) simultaneously spiked into the buffer solution were 87.0 and 99.8%, respectively. Recovery rates of AFB1/DON and DON/ZEA spiked simultaneously were 86.2%/76.6% and 92.0%/86.7%, respectively, at concentrations of 20 ng/mL AFB1, 500 ng/mL ZEA, and 1,000 ng/mL DON. Recoveries using the novel mAb–MNP conjugated system in a buffer solution simultaneously spiked with AFB1, ZEA and DON were 82.5, 94.6 and 73.4%, respectively. Recoveries of DON in animal feed were 107.7–132.5% at concentrations of 250–1,000 ng/g spiked in feed. The immunoaffinity chromatography (IAC) clean-up method was compared with the purification method using novel mAb–MNP. After fortification of animal feed with AFB1 (5, 10 and 20 ng/g feed) and ZEA (125, 250 and 500 ng/g feed), AFB1 and ZEA were purified using both the methods. In the case of the novel mAb-MNP conjugated system, mean recoveries for AFB1 were 89.4, 73.1 and 88.3% at concentrations of 5, 10 and 20 ng/g feed, respectively. For ZEA, mean recoveries were 86.7, 85.9 and 79.1% at concentrations of 125, 250 and 500 ng/g, respectively. For IAC purification, recoveries were 42.9–45.1% for AFB1 and 96.8–103.2% for ZEA. In conclusion, the present purification method using monoclonal antibodies conjugated to MNPs can be used for simple and simultaneous purification of mycotoxins from feed and maize.

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Screening and development of monoclonal antibodies for identification of ferret T follicular helper cells

Scientific Reports ◽

10.1038/s41598-021-81389-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Wenbo Jiang ◽

Julius Wong ◽

Hyon-Xhi Tan ◽

Hannah G. Kelly ◽

Paul G. Whitney ◽

...

Keyword(s):

Animal Model ◽

Monoclonal Antibodies ◽

Lymph Node ◽

Cross Reactivity ◽

Tfh Cells ◽

Follicular Helper ◽

Lymph Node Cells ◽

Human Viruses ◽

Humoral Responses ◽

Murine Monoclonal Antibodies

AbstractThe ferret is a key animal model for investigating the pathogenicity and transmissibility of important human viruses, and for the pre‐clinical assessment of vaccines. However, relatively little is known about the ferret immune system, due in part to a paucity of ferret‐reactive reagents. In particular, T follicular helper (Tfh) cells are critical in the generation of effective humoral responses in humans, mice and other animal models but to date it has not been possible to identify Tfh in ferrets. Here, we describe the screening and development of ferret-reactive BCL6, CXCR5 and PD-1 monoclonal antibodies. We found two commercial anti-BCL6 antibodies (clone K112-91 and clone IG191E/A8) had cross-reactivity with lymph node cells from influenza-infected ferrets. We next developed two murine monoclonal antibodies against ferret CXCR5 (clone feX5-C05) and PD-1 (clone fePD-CL1) using a single B cell PCR-based method. We were able to clearly identify Tfh cells in lymph nodes from influenza infected ferrets using these antibodies. The development of ferret Tfh marker antibodies and the identification of ferret Tfh cells will assist the evaluation of vaccine-induced Tfh responses in the ferret model and the design of novel vaccines against the infection of influenza and other viruses, including SARS-CoV2.

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