Identification of Low Molecular Weight Allergens of American Cockroach and Production of Monoclonal Antibodies

1996 ◽  
Vol 76 (2) ◽  
pp. 195-203 ◽  
Author(s):  
Chii-Huei Wu ◽  
Min-Jium Hsieh ◽  
Jyh-Hsing Huang ◽  
Sheu-Fen Luo
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
American Cockroach ◽  
Low Molecular Weight

Monoclonal antibodies against the capsular K antigen of Escherichia coli (09: K30(A): H12): characterisation and use in analysis of K antigen organisation on the cell surface

1988 ◽  
Vol 34 (10) ◽  
pp. 1159-1165 ◽  
Author(s):  
Mary K. Homonylo ◽  
Sheila J. Wilmot ◽  
Joseph S. Lam ◽  
Leslie A. MacDonald ◽  
Christopher Whitfield
Keyword(s):  
Escherichia Coli ◽  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
Cell Surface ◽  
Molecular Probes ◽  
Low Molecular Weight ◽  
Gel Chromatography ◽  
E Coli ◽  
Capsular Antigen ◽  
K Antigen

Monoclonal antibodies were produced against the capsular antigen of Escherichia coli serotype K(A)30, using a mouse hybridoma system. The antibodies also recognised the chemically identical capsular polysaccharide produced by Klebsiella K20. Chemical modification of the K30 polysaccharide indicated that the glucuronic acid residues found in the E. coli K30 capsular antigen were important in the epitope recognised by these antibodies. Use of the antibodies as molecular probes revealed the presence of two discrete forms of the K30 antigen. One form was comprised of high molecular weight polysaccharide, present as a surface capsular layer. The second form of the antigen was of low molecular weight and was associated with lipopolysaccharide fractions from cell surface polysaccharide extracts. Separation of lipopolysaccharide fractions using gel chromatography in the presence of detergent showed that the low molecular weight K-antigenic fraction comigrated with a lipopolysaccharide lipid A core fraction present in encapsulated E. coli K30 bacteria but absent in acapsular mutants.

Analysis of proteins in rabbit pulmonary surfactant using monoclonal antibodies

1986 ◽  
Vol 250 (3) ◽  
pp. C460-C467 ◽  
Author(s):  
R. J. King ◽  
H. M. Martin ◽  
J. B. Baseman ◽  
J. Morrison-Plummer
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
High Molecular Weight ◽  
Gel Electrophoresis ◽  
Pulmonary Surfactant ◽  
Polyacrylamide Gel Electrophoresis ◽  
Low Molecular Weight ◽  
Weight Group ◽  
Molecular Weights ◽  
Intermediate Group

We have used monoclonal antibodies developed against the apolipoproteins associated with pulmonary surfactant purified from rabbit lavage fluid to study the expression of epitopes common to these proteins. The pulmonary surfactant contained nearly 20 proteins, of which at least 10 were not derived from serum. Electrophoresis, with sulfhydryl reduction of these proteins indicated apparent molecular weights of approximately 155, 135, 125, and 115 X 10(3) (high-molecular-weight group); 80, 70, and 60 X 10(3) (intermediate group); and 18 through 10 X 10(3) (low-molecular-weight group). Two-dimensional polyacrylamide gel electrophoresis, in which the proteins were electrophoresed without reduction in the first dimension, but with sulfhydryl reduction in the second dimension, revealed that the 80, 70, and 60 X 10(3) proteins dissociated into proteins of nominal molecular weights of 40, 35, and 30 X 10(3), respectively. In contrast, the 125 and 115 X 10(3) proteins of the high-molecular-weight group contained a protein which could only be reduced to a minimum molecular weight of 55 to 60 X 10(3). Monoclonal antibodies generally were of three types: those that reacted strongly with the high-molecular-weight group and weakly with the intermediate group; those that reacted conversely; and those that reacted only with the low-molecular-weight group. Our results indicate that at least two different surfactant apolipoproteins, with differing minimum molecular weights in SDS-polyacrylamide gel electrophoresis, have common epitopes. Although these results cannot certify a physiological relationship between these proteins, they suggest that the intracellular synthesis or extracellular processing of surfactant apolipoproteins may be more complicated than predicted by the findings of previous experiments, perhaps involving the posttranslational assembly of one surfactant protein into oligomers which resist dissociation under the conditions used for the analyses.

scholarly journals On-column disulfide bond formation of monoclonal antibodies during Protein A chromatography eliminates low molecular weight species and rescues reduced antibodies

mAbs ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 1829333
Author(s):  
Zhijun Tan ◽  
Vivekh Ehamparanathan ◽  
Tingwei Ren ◽  
Peifeng Tang ◽  
Laurel Hoffman ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
Disulfide Bond ◽  
Bond Formation ◽  
Protein A ◽  
Low Molecular Weight ◽  
Disulfide Bond Formation

Immunolocalization and characterization of the low molecular weight antigen (4–5 kDa) ofToxoplasma gondiithat elicits an early IgM response upon primary infection

Parasitology ◽  
1994 ◽  
Vol 108 (2) ◽  
pp. 139-145 ◽  
Author(s):  
S. Tomavo ◽  
G. Couvreur ◽  
M. A. Leriche ◽  
A. Sadak ◽  
A. Achbarou ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
Primary Infection ◽  
Low Molecular Weight ◽  
Immuno Electron Microscopy ◽  
Surface Localization

SUMMARYA striking feature of toxoplasmic seroconversion is the prominent and early IgM response to a low molecular weight antigen of 4–5 kDa. Two different monoclonal antibodies directed against the 4–5 kDa antigen have been generated and used to characterize this molecule. Using these monoclonal antibodies, we could demonstrate the surface localization of the lowMrantigen by immunofluorescence and immuno-electron microscopy assays. By immunoblotting, we observed that one of the monoclonal antibodies was unable to recognize the 4–5 kDa antigen in tachyzoites propagated in cell culture, indicating an epitope variability betweenToxoplasma gondiitachyzoites grownin vivoandin vitro. We discuss the implications of this latter finding in the design of diagnostic reagents.

Monoclonal Antibodies Selective for Low Molecular Weight Neurofilaments

2002 ◽  
Vol 21 (1) ◽  
pp. 53-59 ◽  
Author(s):  
Niklas Norgren ◽  
Jan-Erik Karlsson ◽  
Lars Rosengren ◽  
Torgny Stigbrand
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
Low Molecular Weight

Insights on Monoclonal Antibodies to Kininogens' Heavy Chain Which Influence Kininogens' Binding to Platelets

1992 ◽  
Vol 68 (02) ◽  
pp. 143-148 ◽  
Author(s):  
Yongping Jiang ◽  
Weerasak Nawarawong ◽  
Frank J Meloni ◽  
Alvin H Schmaier
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
Affinity Chromatography ◽  
Immunosorbent Assay ◽  
Heavy Chain ◽  
Gel Filtration ◽  
Enzyme Linked Immunosorbent Assay ◽  
Low Molecular Weight ◽  
High Concentrations ◽  
Sepharose 4B

SummaryPurified domains of low molecular weight kininogen (LK) can be used directly to determine the epitopes of monoclonal antibodies (mAbs) that have been shown to influence kininogen function. LK, purified from plasma by carboxymethyl-papain-Sepharose 4B affinity chromatography and kaolin adsorption, was digested by trypsin and chymotrypsin. The domains of LK were then separated by gel filtration followed by carboxymethyl-papain-Sepharose 4B affinity chromatography. Using the purified domains of LK’s heavy chain, the regions on kininogens' heavy chain which various monoclonal antibodies are directed to were determined by enzyme-linked immunosorbent assay and immunoblotting. MAb 2B5 which neutralized kininogens' ability to inhibit calpain cross-reacted with domains 2 and 3. MAb HKH8 which reacted with kininogens' domain 1 and 2 was found to inhibit 125I-HK binding to platelets. At two-fold molar excess, mAb HKH8 was a better inhibitor of 125I-HK binding to platelets than higher concentrations, where the antibody was shown to cause increased binding to platelets. Alternatively, HKH8 F(ab')2 completely inhibited 125I-HK binding to platelets even at high concentrations of antibody. These studies indicate that purified domains of kininogens' heavy chain can be used to rapidly localize epitopes for antibodies. Further, mAb HKH8 should be a valuable probe to understand the mechanisms of kininogens' binding to platelets.

Detection of low-molecular-weight polypeptides on nitrocellulose with monoclonal antibodies

1989 ◽  
Vol 183 (2) ◽  
pp. 250-257 ◽  
Author(s):  
Lawrence C. Rosenbaum ◽  
Gajanan Nilaver ◽  
Heidi M. Hagman ◽  
Edward A. Neuwelt
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
Low Molecular Weight

Monoclonal Antibodies Against Human High Molecular Weight Urinary Urokinase: Application for Affinity Purification of Urinary Prourokinase

1986 ◽  
Vol 55 (03) ◽  
pp. 347-351 ◽  
Author(s):  
J Wojta ◽  
J C Kirchheimer ◽  
Liselotte Turcu ◽  
G Christ ◽  
B R Binder
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
High Molecular Weight ◽  
Gel Electrophoresis ◽  
Affinity Purification ◽  
Low Molecular Weight ◽  
Plasminogen Activation ◽  
Single Chain ◽  
Apparent Homogeneity ◽  
Step Procedure

SummaryMonoclonal antibodies against urinary urokinase were obtained by immunizing mice with purified human high molecular weight urokinase. Five antibodies were selected and denominated MPW1UK, MPW2UK, MPW3UK, MPW4UK, and MPW5UK, respectively. All selected antibodies reacted with high and low molecular weight urokinase. Cleavage of the low molecular weight paranitroanilide substrate pyro-Glu-Gly-Arg-pNA by urokinase was not inhibited by the antibodies and only one antibody (MPW5UK) inhibited plasminogen activation by urokinase. The ability of MPW5UK to bind to coated urokinase was 100-fold higher than that of the other antibodies. MPW5UK was used to prepare an immunosorbent for the purification of urokinase antigen from freshly voided crude urine. One-chain prourokinase was separated from two-chain urokinase by chromatography of the urokinase antigen containing mixture on agmatine Sepharose. As judged by SDS gel electrophoresis one-chain prourokinase as well as two-chain urokinase were purified to apparent homogeneity by this two-step procedure; the yields were 18% and 47% for single-chain prourokinase and two-chain urokinase, respectively, as calculated from total urokinase antigen contained in the starting material.

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