Effect of low temperature stress on the expression of sucrose synthetase in spring and winter wheat plants. Development of a monoclonal antibody against wheat germ sucrose synthetase

1991 ◽  
Vol 69 (1) ◽  
pp. 36-41 ◽  
Author(s):  
W. Jay Newsted ◽  
Ravindra N. Chibbar ◽  
Fawzy Georges
Keyword(s):  
Monoclonal Antibody ◽  
Wheat Germ ◽  
Mrna Level ◽  
Cross Reactivity ◽  
Sharp Rise ◽  
Wheat Varieties ◽  
Carrot Cells ◽  
Sucrose Synthetase ◽  
Gradual Accumulation ◽  
Maize Plants

A monoclonal antibody against wheat germ sucrose synthetase is developed and characterized. Its use in studying the effect of cold acclimation on the expression of sucrose synthetase in winter and spring wheat plants is described. The antibody shows cross-reactivity with sucrose synthetase from maize and pea plants, as well as carrot cells. A gradual accumulation of the enzyme as a function of time spent at 2 °C is observed in both wheat varieties. In contrast, an initial sharp rise in the mRNA level is observed, which agrees with the previously reported response of maize plants subjected to anaerobic stress.Key words: cold tolerance, antibodies, protein expression, sucrose synthetase.

Evidence of Cross-Reactivity of ‘Carcinoma-Specific’ KC4 Monoclonal Antibody with Activated Mesothelial Cells and Phytohemagglutinin-Stimulated Lymphocytes

Oncology ◽  
1988 ◽  
Vol 45 (5) ◽  
pp. 380-383
Author(s):  
A. Neubauer ◽  
R. Musch ◽  
U. Thalmann ◽  
H. Grosser ◽  
J. Laser ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Cross Reactivity ◽  
Mesothelial Cells

Hevein-like domain IV in wheat germ agglutinin (WGA) is responsible for the majority of IgE cross-reactivity between WGA and hevein (Hev b 6.02)

2003 ◽  
Vol 111 (2) ◽  
pp. S327
Author(s):  
P. Karisola ◽  
H. Alenius ◽  
K. Turjanmaa ◽  
T. Reunala ◽  
N. Kalkkinen ◽  
...  
Keyword(s):  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Cross Reactivity

scholarly journals Generation of a Monoclonal Antibody against D-Dimer Using HTS-Based LiCA

2019 ◽  
Vol 25 (3) ◽  
pp. 310-319
Author(s):  
Yuan Dong ◽  
Hanjin Hou ◽  
An Chen ◽  
Wei Ma ◽  
Moli Yin ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Degradation Products ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cross Reactivity ◽  
Enzyme Hydrolysis ◽  
Clinical Samples ◽  
Sds Page ◽  
Thrombotic Diseases ◽  
D Dimer

D-dimer is an essential diagnostic index of thrombotic diseases. Since the existing anti-D-dimer antibodies vary in quality and specificity, a search for alternative anti-D-dimer antibodies is required. The present study aimed to screen a novel monoclonal antibody (mAb) against D-dimer using a light-initiated chemiluminescence assay (LiCA). In this work, mice were immunized with antigen prepared from human plasma by enzyme hydrolysis. After screening, a novel mAb, DD 2G11, was obtained. The results of sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis indicated that DD 2G11 could be used as a standard marker for D-dimer. The isotype of DD 2G11 was IgG1, the Ka value was 0.646 nM-1, and the Kd value was 50 nM, indicating that the binding affinity to D-dimer was very high. Furthermore, no cross-reactivity between DD 2G11 and other fibrinogen degradation products (FgDPs) was found. Finally, the correlation between DD 2G11 and the reference antibody (commercial antibody) was investigated by analyzing 56 clinical samples using a latex-enhanced turbidimetric immunoassay (LTIA). The R2 value of the linear regression was 0.94538, indicating that DD 2G11 met clinical requirements. In conclusion, the present study provides a more expeditious protocol to screen mAbs and provides a clinically usable mAb against D-dimer.

A validation study of selected methods routinely used for measurement of cyclosporine

1990 ◽  
Vol 36 (4) ◽  
pp. 670-674 ◽  
Author(s):  
K E Blick ◽  
S H Melouk ◽  
H D Fry ◽  
R L Gillum
Keyword(s):  
Monoclonal Antibody ◽  
Polyclonal Antibody ◽  
Validation Study ◽  
Liver Dysfunction ◽  
Correlation Coefficients ◽  
Cross Reactivity ◽  
Poor Correlation ◽  
Fluorescence Polarization Immunoassay ◽  
Gamma Glutamyltransferase ◽  
Transplant Patients

Abstract We compare four methods for measuring cyclosporine (CyA) in plasma and whole blood of transplant patients: HPLC, RIA with a polyclonal antibody, RIA with a monoclonal antibody, and fluorescence polarization immunoassay (FPIA). The monoclonal RIA procedure correlated acceptably with HPLC, with slope = 1.21, r = 0.97, and Sy,x = +/- 40.1. However, the FPIA, done in three separate instruments, correlated relatively poorly with HPLC, giving slopes of 1.67, 1.51, and 2.32; correlation coefficients of 0.72, 0.43, and 0.83; and Sy,x = +/- 205.4, +/- 334.5, and +/- 222.4. The polyclonal RIA correlated reasonably well with HPLC, with a slope = 1.15, r = 0.90, and Sy,x = +/- 72.6. Values for individual patients with increases both in gamma-glutamyltransferase and creatinine showed very poor correlation between FPIA and HPLC, which suggests that metabolite cross-reactivity with FPIA is significant and unpredictable in patients with liver dysfunction coexisting with renal dysfunction. Evidently, the monoclonal RIA can be substituted for HPLC, if the therapeutic range is adjusted for the 21% higher results obtained by RIA.

scholarly journals Structure-Based Modification of an Anti-neuraminidase Human Antibody Restores Protection Efficacy against the Drifted Influenza Virus

mBio ◽  
2020 ◽  
Vol 11 (5) ◽  
Author(s):  
Haihai Jiang ◽  
Weiyu Peng ◽  
Jianxun Qi ◽  
Yan Chai ◽  
Hao Song ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Influenza Virus ◽  
Antibody Fragment ◽  
Salt Bridge ◽  
Cross Reactivity ◽  
Human Antibody ◽  
Influenza Strain ◽  
Group 2 ◽  
Fragment Antigen Binding ◽  
Group 1

ABSTRACT Here, we investigate a monoclonal antibody, Z2B3, isolated from an H7N9-infected patient, that exhibited cross-reactivity to both N9 (group 2) and a broad range of seasonal and avian N1 (group 1) proteins but lost activity to the N1 with the substitution K432E. This substitution exists in 99.25% of seasonal influenza strains after 2013. The NA-Z2B3 complex structures indicated that Z2B3 binds within the conserved active site of the neuraminidase (NA) protein. A salt bridge between D102 in Z2B3 and K432 in NA plays an important role in binding. Structure-based modification of Z2B3 with D102R in heavy chain reversed the salt bridge and restored the binding and inhibition of N1 with E432. Furthermore, Z2B3-D102R can protect mice from A/Serbia/NS-601/2014 H1N1 virus (NA contains E432) infection while the wild-type Z2B3 antibody shows no protection. This study demonstrates that a broadly reactive and protective antibody to NA can be in principle edited to restore binding and inhibition to recently drifted N1 NA and regain protection against the variant influenza strain. IMPORTANCE The immune system produces antibodies to protect the human body from harmful invaders. The monoclonal antibody (MAb) is one kind of effective antivirals. In this study, we isolated an antibody (Z2B3) from an H7N9 influenza virus-infected child. It shows cross-reactivity to both group 1 (N1) and group 2 (N9) neuraminidases (NAs) but is sensitive to N1 NA with a K432E substitution. Structural analysis of the NA-antibody fragment antigen-binding (Fab) complex provides a clue for antibody modification, and the modified antibody restored binding and inhibition to recently drifted N1 NA and regained protection against the variant influenza strain. This finding suggests that antibodies to NA may be a useful therapy and can be in principle edited to defeat drifted influenza virus.

Listeriolysin O-Based Latex Agglutination Test for the Rapid Detection of Listeria monocytogenes in Foods

1997 ◽  
Vol 60 (9) ◽  
pp. 1038-1040 ◽  
Author(s):  
GHASSAN M. MATAR ◽  
PEGGY S. HAYES ◽  
WILLIAM F. BIBB ◽  
BALA SWAMINATHAN
Keyword(s):  
Monoclonal Antibody ◽  
Listeria Monocytogenes ◽  
Rapid Detection ◽  
Rapid Test ◽  
Food Samples ◽  
Cross Reactivity ◽  
Latex Agglutination ◽  
Listeriolysin O ◽  
Agglutination Assay ◽  
Latex Beads

A latex agglutination-based test for the rapid detection of Listeria monocytogenes in foods was developed. An antilisteriolysin O (LLO) monoclonal antibody (HID5E12D7; IgG2b) covalently bound to polystyrene amidine-modified latex beads was used in a slide agglutination assay. The latex reagent detected 0.1 ng/ml of LLO in phosphate-buffered saline plus bovine serum albumin. It reacted with culture supernatants of L. monocytogenes but not with other Listeria species or Streptococcus groups A through G. The listeriolysin O latex agglutination assay (LLOLAT) was applied to 24-h and 48-h USDA primary enrichment cultures of 208 food samples obtained from refrigerators of listeriosis patients enrolled in a study to determine the role of foods in sporadic listeriosis. Of 19 samples positive by cultural techniques, 17 were positive by the LLOLAT. Cultures with low (<0.3 CFU/g) levels of L. monocytogenes were positive in the LLOLAT. No cross-reactivity occurred when using a heterogeneous monoclonal antibody. The LLOLAT is a sensitive, specific and rapid test and may be useful for screening foods for L. monocytogenes.

scholarly journals Immunoadsorption assay for bone alkaline phosphatase compared with wheat germ agglutinin precipitation assay in serum from (pre)pubertal girls

1996 ◽  
Vol 42 (12) ◽  
pp. 1970-1974 ◽  
Author(s):  
A A Bouman ◽  
C M de Ridder ◽  
J H Nijhof ◽  
J C Netelenbos ◽  
H A Delemarre-vd Waal
Keyword(s):  
Alkaline Phosphatase ◽  
Correlation Coefficient ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Bone Alkaline Phosphatase ◽  
Cross Reactivity ◽  
Reference Ranges ◽  
Correlation Studies ◽  
Deming Regression ◽  
The Mean

Abstract The performance characteristics of two bone alkaline phosphatase (ALP; EC 3.1.3.1) assays, a wheat germ agglutinin (WGA) precipitation assay and a new immunoadsorption assay (IAA), were compared. The within- and between-run imprecision of the IAA (3.6-4.2% and 3.6-7.7%) was comparable with that of the WGA assay. The mean cross-reactivity with liver ALP appeared to be 4% in the WGA assay and 11% in the IAA. The reference ranges in a group of 155 healthy Caucasian (pre)pubertal schoolgirls were: 149-401 U/L (total ALP, 30 degrees C), 105-349 U/L (bone ALP, 30 degrees C, WGA assay), and 58-205 U/L (bone ALP, 25 degrees C, IAA). Comparison of the WGA assay (x) with the IAA (y) demonstrated a correlation coefficient of 0.95 [Deming regression equation: y = (0.56 +/- 0.01)x + (2.0 +/- 1.5); Sy[symbol: see text]x = 5.3 U/L]. Correlation studies of the WGA assay and the IAA results with total ALP demonstrated r = 0.98 and 0.96, respectively.

Molecular characterization of a partial sequence encoding a novel Schistosoma mansoni serine protease

Parasitology ◽  
1997 ◽  
Vol 115 (4) ◽  
pp. 395-402 ◽  
Author(s):  
C. COCUDE ◽  
C. PIERROT ◽  
C. CETRE ◽  
C. GODIN ◽  
A. CAPRON ◽  
...  
Keyword(s):  
Schistosoma Mansoni ◽  
Serine Protease ◽  
Serine Proteases ◽  
Mrna Level ◽  
Cross Reactivity ◽  
Reading Frame ◽  
Consensus Sequences ◽  
Degenerate Oligonucleotide ◽  
Sequence Encoding

A PCR strategy using degenerate oligonucleotide primers based upon consensus sequences of the active site of serine proteases yielded a 467 bp fragment from genomic DNA from Schistosoma mansoni cercariae. The sequence presented a continuous open reading frame and the deduced amino acid sequence (156 aa) presented homologies with various serine proteases, in particular the highest percentage identity was observed with a mammalian plasma kallikrein. The expression of this serine protease was studied first at the mRNA level and it was only detected by RT-PCR in cercariae and in adult worms. At the protein level we were able to detect it by Western blotting and by using antigen extracts from metabolically radio-isotope labelled worms. The absence of any positive signal in Northern blot and the detection of the protein suggest that the mRNA has a very short half-life, however the protein may be accumulated in the parasite. The significance of identity with mammalian kallikrein was confirmed by cross-immunoreactivity with a native porcine pancreatic kallikrein. However, no cross-reactivity was observed with S. mansoni elastase, another serine protease. Thus, we suggest that the serine protease described in this paper is a kallikrein-like protease.

scholarly journals A NS1-binding monoclonal antibody interacts with two residues that are highly conserved in seasonal as well as newly emerged influenza A virus

2019 ◽  
Vol 77 (1) ◽  
Author(s):  
Su Hui Catherine Teo ◽  
Jian-Ping Wu ◽  
Chee-Keng Mok ◽  
Yee-Joo Tan
Keyword(s):  
Monoclonal Antibody ◽  
Influenza A Virus ◽  
Influenza A ◽  
Rna Binding ◽  
Intracellular Localization ◽  
Cross Reactivity ◽  
Structural Protein ◽  
Multifunctional Protein ◽  
Good Target ◽  
Conformational Plasticity

Abstract The non-structural protein 1 (NS1) of influenza A virus (IAV) is a multifunctional protein that antagonizes host antiviral responses, modulating virus pathogenesis. As such, it serves as a good target for research and diagnostic assay development. In this study, we have generated a novel monoclonal antibody (mAb) 19H9 and epitope mapping revealed that two residues, P85 and Y89, of NS1 are essential for interacting with this mAb. Furthermore, residues P85 and Y89 are found to be highly conserved across different IAV subtypes, namely seasonal H1N1 and H3N2, as well as the highly pathogenic H5N1 and H5N6 avian strains. Indeed, mAb 19H9 exhibits broad cross-reactivity with IAV strains of different subtypes. The binding of mAb 19H9 to residue Y89 was further confirmed by the abrogation of interaction between NS1 and p85β. Additionally, mAb 19H9 also detected NS1 proteins expressed in IAV-infected cells, showing NS1 intracellular localization in the cytoplasm and nucleolus. To our knowledge, mAb 19H9 is the first murine mAb to bind at the juxtaposition between the N-terminal RNA-binding domain and C-terminal effector domain of NS1. It could serve as a useful research tool for studying the conformational plasticity and dynamic changes in NS1.

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