Effect of Heat Treatment on the Antigen-Binding Activity of Anti-Peroxidase Immunoglobulins in Bovine Colostrum

Abstract

After intramammary immunization with horseradish peroxidase, bovine colostrum containing anti-peroxidase immunoglobulins (Ig) was obtained. Thermoresistance of the antigen-binding region of these Ig was studied using a direct competitive ELISA. This technique is based on the competition between the anti-peroxidase IgG coated to the plate and the anti-peroxidase IgG contained in the colostrum to bind peroxidase, the antigen, and the enzyme responsible for development of the color of the assay. Thus, the degree of denaturation of the antigen-binding region in the IgG molecule can be determined because this region is directly involved in the assay. The kinetic and thermodynamic parameters for heat induced denaturation of IgG in colostrum were determined over a temperature range of 69 to 81 degrees C. The denaturation of IgG was best described assuming an apparent reaction order of 1.5. D values, the time required to reduce the antigen-binding activity of IgG by 90%, were 8504, 1387, 285, and 152 s at 69, 72, 77, and 81 degrees C, respectively. Similarly, Z value, the degrees necessary to reduce the D value in one logarithmic cycle, was estimated to be 6.6 degrees C, and the activation energy value was 386.83 kJ/mol. These results should be taken into account in the design of heat treatments of milk in order to preserve the biological function of Ig.

Analysis of denaturation of bovine IgG by heat and high pressure using an optical biosensor

“…Other studies of the thermal denaturation of bovine milk IgG have variously reported reaction orders between 1.2 and 2.5, with fractional orders indicating multiple simultaneous unfolding and aggregation pathways (Cao et al, 2007;Dominguez et al, 1997;Law, 1995;Levieux, Levieux, El-Hatmi, & Rigaudiere, 2006;Mainer et al, 1997). It is difficult to rationalise the discrepancies in reported denaturation reaction order under comparable conditions, although the dependency of the rate constant on initial concentration and on temperature, where unfolding and aggregation may be rate-determining over different ranges, will be contributing factors.…”

“…However, few studies have considered differentiation of Fab and Fc domains within bovine lacteal IgG with respect to relative conformational stability, and the potential for other milk components to moderate this stability, and conclusions remain equivocal (Cao et al, 2007;Dominguez et al, 1997Dominguez et al, , 2001Li et al, 2005;Ohnuki & Otani, 2005). A study of the stability of mouse monoclonal IgG by calorimetric and circular dichroism techniques has revealed independent denaturation transitions of Fab and Fc domains, with each region being influenced distinctively by thermal or pH stress (Vermeer & Norde, 2000).…”

“…Several studies have reported on the stability of bovine IgG, either in model systems or in intact milk or colostrum, to various stresses including storage, isolation and processing variables such as heat, pH and pressure (Ando et al, 2005;Cao, Wang, & Zheng, 2007;Chen & Chang, 1998;Chen, Tu, & Chang, 2000;Dominguez, Pe´rez, & Calvo, 1997;Dominguez, Pe´rez, Puyol, Sa´nchez, & Calvo, 2001;Law, 1995;Li, Bomser, & Zhang, 2005;Li et al, 2006;Li-Chan, Kummer, Losso, Kitts, & Nakai, 1995;Lindstro¨m, Paulsson, Nylander, Elofsson, & Lindmark-Ma˚nsson, 1994; Mainer, Sa´nchez, Ena, & Calvo, 1997;Ohnuki & Otani, 2005;Rosenthal, Bernstein, & Merin, 1999). These studies have utilised a range of spectroscopic, immunoassay and separation-based analytical techniques and have inevitably reported widely variable stability and reaction order estimates.…”

Electrospray–differential mobility analysis of bionanoparticles

“…For n ≠ 1, the general rate equation can be solved to get: [207], 1.5 [208,209], 2.0 [210,211] or 2.5 [212]. The order of the reaction is typically determined by fitting the experimental data using different reaction orders, and assigning the order with highest R 2 value.…”

Alimentación y manejo del calostro en el ganado de leche

“…Algunos de los patógenos que pueden estar presentes en el calostro, ya sea provenientes de la glándula mamaria o de la contaminación en el manejo del mismo y que pueden ser transmitidos a las terneras incluyen: Mycobacterium avium spp. Paratuberculosis, Salmonella spp., Micoplasma spp., Listeria monocytogens Campylobacter spp., Mycobacterium bovis y Escherichia coli (Dominguez 1997;Stabel 2001;Stabel 2004;Stewart 2005;Godden et al 2006). Estos agentes infecciosos pueden ocasionar enfermedades como la enteritis y septicemia.…”