Rabbit antibodies against the low molecular weight folate binding protein from human milk. Use for immunological characterization of human folate binding proteins in an enzyme-linked immunosorbent assay (ELISA)

1987 ◽  
Vol 7 (7) ◽  
pp. 553-557 ◽  
Author(s):  
Mimi Høier-Madsen ◽  
Steen Ingemann Hansen ◽  
Jan Holm
Keyword(s):  
Molecular Weight ◽  
Human Milk ◽  
Immunosorbent Assay ◽  
Binding Proteins ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Low Molecular Weight ◽  
Folate Binding Protein ◽  
Folate Binding Proteins

Antibodies to a low molecular weight folate binding protein isolated from human milk were raised in rabbits and used for development of a two-site enzyme-linked immunosorbent assay (ELISA) for immunological characterization of human folate binding proteins (FBPs). The high and low molecular weight FBPs from human milk were immunologically indistinguishable. Furthermore, the FBPs in human urine and cerebrospinal fluid showed a cross-reactivity of 70% and 30%, respectively. No cross-reactivity of the FBP from cow's milk was observed.

Rabbit antibodies against the folate binding protein from cow's milk. Production, characterization and use for development of an enzyme-linked immunosorbent assay (ELISA)

1986 ◽  
Vol 6 (10) ◽  
pp. 895-900 ◽  
Author(s):  
Mimi Høier-Madsen ◽  
Steen Ingemann Hansen ◽  
Jan Holm
Keyword(s):  
Immunosorbent Assay ◽  
Binding Proteins ◽  
Binding Protein ◽  
Cross Reactivity ◽  
Cow’S Milk ◽  
Enzyme Linked Immunosorbent Assay ◽  
Folate Binding Protein ◽  
Milk Whey ◽  
Cow's Milk ◽  
Folate Binding Proteins

Rabbit antibodies to purified folate binding protein from cow's milk whey were used for development of a two-site enzyme-linked immunosorbent assay (ELISA) for quantitation of bovine folate binding proteins, The folate binding proteins in human milk and serum showed no cross-reactivity. A partial saturation of purified bovine folate binder with folate gave rise to an increased antigenicity probably due to a ligand (folate)-induced exposure of antigenic sites on the protein.

High-affinity folate binding in human prostate

1993 ◽  
Vol 13 (2) ◽  
pp. 99-105 ◽  
Author(s):  
Jan Holm ◽  
Steen Ingemann Hansen ◽  
Mimi Høier-Madsen
Keyword(s):  
Human Milk ◽  
Binding Protein ◽  
Cross Reactivity ◽  
Human Prostate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Gel Chromatography ◽  
Folate Binding Protein ◽  
High Affinity ◽  
Sds Page ◽  
Triton X

Binding of 3H-folate in Triton X-100 solubilized human prostate homogenate was of a high-affinity type and displayed apparent positive cooperativity typical of specific folate binding. Radioligand dissociation was slow at pH 7.4, but rapid at pH 3.5. Gel chromatography reveled two major folate binding proteins (Mr≈100 and 25kDa), but only one single band (Mr ≈ 65–70 kDa) was detectable on SDS-PAGE and immunoblotting with rabbit-anti human milk folate binding protein. Concentration of folate binding protein in prostate homogenate expressed as maximum 3H-folate binding was 1.10 nmol/g protein, and the cross-reactivity with rabbit-anti human milk folate binding protein serum was 15% as determined by an enzyme-linked immunosorbent assay (median values; n = 6).

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.

Immunological analysis ofOpisthorchisandClonorchisantigens

1990 ◽  
Vol 64 (2) ◽  
pp. 133-138 ◽  
Author(s):  
S. Sirisinha ◽  
D. Sahassananda ◽  
D. Bunnag ◽  
H. J. Rim
Keyword(s):  
Molecular Weight ◽  
Immunosorbent Assay ◽  
Clonorchis Sinensis ◽  
Cross Reactivity ◽  
Opisthorchis Viverrini ◽  
Metabolic Product ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immunological Analysis ◽  
Metabolic Products ◽  
Liver Flukes

ABSTRACTImmunoreactive components ofOpisthorchis viverriniandClonorchis sinensiswere analysed by enzyme-linked immunosorbent assay (ELISA), radioimmunoprecipitation and immunoblotting. Somatic extracts from these two liver flukes as well as from other related parasites, together with the metabolic products, were tested for their reactivities with sera from patients with opisthorchiasis and clonorchiasis. A significant cross-reactivity in the ELISA was noted betweenOpisthorchisandClonorchis. Immunoblotting and radioimmunoprecipitation analyses showed that the 89-kD protein which was previously shown to be a predominant metabolic product ofO. viverrinireacted with sera from both groups of patients. However, an antigen with a molecular weight of 16 kD, apparently a predominant somatic component, appeared to be specific forO. viverrini.

Immunochemical Quantification of Metallothioneins of a Marine Mollusc

1988 ◽  
Vol 45 (7) ◽  
pp. 1257-1263 ◽  
Author(s):  
G. Roesijadi ◽  
M. E. Unger ◽  
J. E. Morris
Keyword(s):  
Metal Binding ◽  
Binding Proteins ◽  
Polyclonal Antibodies ◽  
Exchange Chromatography ◽  
Enzyme Linked Immunosorbent Assay ◽  
Low Molecular Weight ◽  
Ammonium Sulfate Precipitation ◽  
Marine Mollusc ◽  
Peroxidase Conjugate

Mercury-induced, low molecular weight, metal-binding proteins were isolated from the marine mussel Mytilus edulis and used as antigen in the development of antibodies and an enzyme-linked immunosorbent assay (ELISA) for quantification of the proteins. Partial characterization of the isolated protein indicated that its properties were consistent with those of metallothionein (MT). In contrast with most MTs, this protein occurred as an apparent dimer and contained glycine at high levels. Polyclonal antibodies against this protein were produced in goats and purified to an IgG fraction by ammonium sulfate precipitation and DEAE ion-exchange chromatography. Ouchterlony analysis and ELISA showed that these antibodies were cross-reactive with two other charge variants of mercury-induced, metal-binding proteins of M. edulis, but not with rabbit MT. The ELISA was based on an indirect, competitive procedure utilizing a rabbit anti-goat IgG–horseradish peroxidase conjugate as the second antibody. Application of this assay to cytosolic extracts of mussel gills indicated 0.5 μg metal-binding proteins/g wet tissue weight in gills of control mussels and elevated levels up to 1780 μg/g following exposure to mercury, cadmium, or copper for 28 d. Zinc was not effective as an inducer of these proteins.

CHARACTERIZATION OF THE PIGMENT OF MICROCOCCUS VIOLAGABRIELLAE

1964 ◽  
Vol 10 (5) ◽  
pp. 659-675 ◽  
Author(s):  
J. N. Campbell ◽  
J. L. Nichols ◽  
Sheila A. Berry
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Amino Acid ◽  
Cross Reactivity ◽  
Low Molecular Weight ◽  
Terminal Amino Acid ◽  
Iron Chelate ◽  
Peptide Moiety ◽  
Terminal Amino

Production of the red insoluble pigment by Micrococcus violagabriellae was studied. Pigmentation was found to require oxygen and high levels of iron and to be stimulated by tryptophan alone among the amino acids. The pigment was isolated, purified, and analyzed chemically and spectrophotometrically. It was found to be similar to pulcherrimin from Candida pulcherrima. Immunological cross reactivity and analysis of derivatives confirmed the similarity between the bacterial and yeast pigments. From these data it is postulated that the pigment is an iron chelate of pulcherriminic acid with an associated low molecular weight peptide moiety with glycine as the sole N-terminal amino acid. The pigment appears to differ from that of C. pulcherrima solely with respect to this peptide and in the mode of aggregation of the molecule.

Rap1b Association with the Platelet Cytoskeleton Occurs in the Absence of Glycoproteins IIb/IIIa

1998 ◽  
Vol 79 (04) ◽  
pp. 832-836 ◽  
Author(s):  
Thomas Fischer ◽  
Christina Duffy ◽  
Gilbert White
Keyword(s):  
Molecular Weight ◽  
Divalent Cation ◽  
Binding Proteins ◽  
Binding Protein ◽  
Platelet Membrane ◽  
Low Molecular Weight ◽  
Membrane Glycoproteins ◽  
Gtp Binding Protein ◽  
Gtp Binding Proteins ◽  
Gtp Binding

SummaryPlatelet membrane glycoproteins (GP) IIb/IIIa and rap1b, a 21 kDa GTP binding protein, associate with the triton-insoluble, activation-dependent platelet cytoskeleton with similar rates and divalent cation requirement. To examine the possibility that GPIIb/IIIa was required for rap1b association with the cytoskeleton, experiments were performed to determine if the two proteins were linked under various conditions. Chromatography of lysates from resting platelets on Sephacryl S-300 showed that GPIIb/IIIa and rap1b were well separated and distinct proteins. Immunoprecipitation of GPIIb/IIIa from lysates of resting platelets did not produce rap1b or other low molecular weight GTP binding proteins and immunoprecipitation of rap1b from lysates of resting platelets did not produce GPIIb/IIIa. Finally, rap1b was associated with the activation-dependent cytoskeleton of platelets from a patient with Glanzmann’s thrombasthenia who lacks surface expressed glycoproteins IIb and IIIa. Based on these findings, we conclude that no association between GPIIb/IIIa and rap1b is found in resting platelets and that rap1b association with the activation-dependent cytoskeleton is at least partly independent of GPIIb/IIIa.

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