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.

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.

Indirect enzyme‐linked Immunosorbent assay for detection of cow's milk in goat's milk

1992 ◽  
Vol 4 (1) ◽  
pp. 11-18 ◽  
Author(s):  
C. Castro ◽  
R. Martín ◽  
T. García ◽  
E. Rodríguez ◽  
I. González ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Cow’S Milk ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cow's Milk ◽  
Goat’S Milk ◽  
Goat's Milk

scholarly journals Properties of Folate Binding Protein Purified from Cow's Milk

2012 ◽  
Vol 19 (3) ◽  
pp. 105-109
Author(s):  
A SUBANDRATE ◽  
DWIRINI RETNO GUNARTI ◽  
MOHAMAD SADIKIN
Keyword(s):  
Binding Protein ◽  
Cow’S Milk ◽  
Folate Binding Protein ◽  
Cow's Milk

Binding of Radiolabeled Folate and 5-Methyltetrahydrofolate to Cow's Milk Folate Binding Protein at pH 7.4 and 5.0. Relationship to Concentration and Polymerization Equilibrium of the Purified Protein

2001 ◽  
Vol 21 (6) ◽  
pp. 733-743 ◽  
Author(s):  
Jan Holm ◽  
Steen Ingemann Hansen
Keyword(s):  
Ligand Binding ◽  
Binding Affinity ◽  
Conformational Changes ◽  
Protein Conformation ◽  
Binding Protein ◽  
Cow’S Milk ◽  
Folate Binding Protein ◽  
Cow's Milk ◽  
Polymerization Equilibrium ◽  
Methyl Tetrahydrofolate

Binding of folate (pteroylglutamate) and 5-methyltetrahydrofolate, the major endogenous form of folate, to folate binding protein purified from cow's milk was studied at 7°C to avoid degradation of 5-methyltetrahydrofolate. Both folates dissociate rapidly from the protein at pH 3.5, but extremely slowly at pH 7.4, most likely due to drastic changes in protein conformation occurring after folate binding. Dissociation of 5-methyltetrahydrofolate showed no increase at 37°C suggesting that protein-bound-5-methyltetrahydrofolate is protected against degradation. Binding displayed two characteristics, positive cooperativity and a binding affinity that increased with decreasing concentrations of the protein. The binding affinity of folate was somewhat greater than that of 5-methyl tetrahydrofolate, in particular at pH 5.0. Ligand-bound protein exhibited concentration-dependent polymerization (8-mers formed at 13 μM) at pH 7.4. At pH 5.0, only folate-bound forms showed noticeable polymerization. The fact that folate at pH 5.0 surpasses 5-methyltetrahydrofolate both with regard to binding affinity and ability to induce polymerization suggests that ligand binding is associated with conformational changes of the protein which favor polymerization.

Ionic Charge, Hydrophobicity and Tryptophan Fluorescence of the Folate Binding Protein Isolated from Cow's Milk

2001 ◽  
Vol 21 (3) ◽  
pp. 305-313 ◽  
Author(s):  
Jan Holm ◽  
Steen Ingemann Hansen ◽  
Mimi Høier-Madsen
Keyword(s):  
Ligand Binding ◽  
Binding Protein ◽  
Tryptophan Fluorescence ◽  
Exchange Chromatography ◽  
Cow’S Milk ◽  
Conformation Change ◽  
Ionic Charge ◽  
Folate Binding Protein ◽  
Cow's Milk ◽  
Isoelectric Points

A high-affinity folate binding protein was isolated and purified from cow's milk by a combination of cation exchange chromatography and methotrexate affinity chromatography. Chromatofocusing studies revealed that the protein possessed isoelectric points in the pH-interval 8–7. Polymers of the protein prevailing at pH values close to the isoelectric points seemed to be more hydrophobic than monomers present at pH 5.0 as evidenced by hydrophobic interaction chromatography and turbidity (absorbance at 340 nm) in aqueous buffer solutions (pH 5–8). Ligand binding seemed to induce a conformation change that decreased the hydrophobicity of the protein. In addition, Ligand binding quenched the tryptophan fluorescence of folate binding protein suggesting that tryptophan is present at the binding site and/or ligand binding induces a conformation change that affects tryptophan environment in the protein. There was a noticeable discordance between the ability of individual folate analogues to compete with folate for binding and the quenching effect.

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).

Effect of Methotrexate on Folate Binding to a Folate Binding Protein in Cow's Milk

2009 ◽  
Vol 42 (1) ◽  
pp. 77-80 ◽  
Author(s):  
Jan Holm ◽  
Steen Ingemann Hansen ◽  
Jørgen Lyngbye
Keyword(s):  
Binding Protein ◽  
Cow’S Milk ◽  
Folate Binding Protein ◽  
Cow's Milk

Effect of Hydrogen Ion Concentration and Buffer Composition on Ligand Binding Characteristics and Polymerization of Cow's Milk Folate Binding Protein

2001 ◽  
Vol 21 (6) ◽  
pp. 745-753 ◽  
Author(s):  
Jan Holm ◽  
Steen Ingemann Hansen
Keyword(s):  
Ligand Binding ◽  
Binding Protein ◽  
Hydrogen Ion ◽  
Ion Concentration ◽  
Cow’S Milk ◽  
Folate Binding Protein ◽  
Hydrogen Ion Concentration ◽  
Binding Characteristics ◽  
Cow's Milk ◽  
Buffer Composition

The ligand binding and aggregation behavior of cow's milk folate binding protein depends on hydrogen ion concentration and buffer composition. At pH 5.0, the protein polymerizes in Tris-HCl subsequent to ligand binding. No polymerization occurs in acetate, and binding is markedly weaker in acetate or citrate buffers as compared to Tris-HCl. Polymerization of ligand-bound protein was far more pronounced at pH 7.4 as compared to pH 5.0 regardless of buffer composition. Binding affinity increased with decreasing concentration of protein both at pH 7.4 and 5.0. At pH 5.0 this effect seemed to level off at a protein concentration of 10−6 M which is 100–1000 fold higher than at pH 7.4. The data can be interpreted in terms of complex models for ligand binding systems polymerizing both in the absence or presence of ligand (pH 7.4) as well as only subsequent to ligand binding (pH 5.0).

scholarly journals Comparison of the Allergenicity and Immunogenicity of Camel and Cow’s Milk—A Study in Brown Norway Rats

Nutrients ◽  
2018 ◽  
Vol 10 (12) ◽  
pp. 1903 ◽  
Author(s):  
Natalia Maryniak ◽  
Egon Hansen ◽  
Anne-Sofie Ballegaard ◽  
Ana Sancho ◽  
Katrine Bøgh
Keyword(s):  
Protein Composition ◽  
Cross Reactivity ◽  
Camel Milk ◽  
Brown Norway ◽  
Cow’S Milk ◽  
Infant Formulas ◽  
Promising Alternative ◽  
Norway Rat ◽  
Milk Whey ◽  
Cow's Milk

Background: When breastfeeding is impossible or insufficient, the use of cow’s milk-based hypoallergenic infant formulas is an option for infants suffering from or at risk of developing cow’s milk allergy. As the Camelidae family has a large evolutionary distance to the Bovidae family and as camel milk differs from cow’s milk protein composition, there is a growing interest in investigating the suitability of camel milk as an alternative to cow’s milk-based hypoallergenic infant formulas. Methods: The aim of the study was to compare the allergenicity and immunogenicity of camel and cow’s milk as well as investigating their cross-reactivity using a Brown Norway rat model. Rats were immunised intraperitoneally with one of four products: camel milk, cow’s milk, cow’s milk casein or cow’s milk whey fraction. Immunogenicity, sensitising capacity, antibody avidity and cross-reactivity were evaluated by means of different ELISAs. The eliciting capacity was evaluated by an ear swelling test. Results: Camel and cow’s milk showed similarity in their inherent immunogenicity, sensitising and eliciting capacity. Results show that there was a lower cross-reactivity between caseins than between whey proteins from camel and cow’s milk. Conclusions: The study showed that camel and cow’s milk have a low cross-reactivity, indicating a low protein similarity. Results demonstrate that camel milk could be a promising alternative to cow’s milk-based hypoallergenic infant formulas.

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