Lactoferrin quantification in cattle faeces by ELISA

Abstract

Background: Promoting and maintaining health is critical to ruminant welfare and productivity. Within human medicine, faecal lactoferrin is quantified for routine assessment of various gastrointestinal illnesses avoiding the need for blood sampling. This approach might also be adapted and applied for non-invasive health assessments in animals. Methods: In this proof-of-concept study, a bovine lactoferrin enzyme-linked immunosorbent assays (ELISA), designed for serum and milk, was applied to a faecal supernatant to assess its potential for quantifying lactoferrin in the faeces of cattle. Faecal lactoferrin concentrations were compared to background levels to assess the viability of the technique. A comparison was then made against serum lactoferrin levels to determine if they were or were not reflective of one another. Results: The optical densities of faecal samples were significantly greater than background readings, supporting the hypothesis that the assay was effective in quantifying faecal lactoferrin (T 13, 115 = 11.99, p < 0.0005). The mean faecal lactoferrin concentration was 0.269 µg mL −1 (S.E. 0.031) and the mean serum concentration 0.074 µg mL −1 (S.E. 0.005). Lactoferrin concentrations of faecal and serum samples, taken from the same animals on the same day, were significantly different (T 21 = 2.20, p = 0.039) and did not correlate (r = 0.2699, p = 0.238). Conclusion: Results support the hypothesis that lactoferrin can be quantified in cattle faeces by ELISA. Whilst further research is required to determine the physiological source of the lactoferrin, this highlights the potential of the method for non-invasive assessment of cattle immunology and pathology.

A picogram BA-ELISA quantification assay for rLj-RGD3, a platelet fibrinogen receptor antagonist, in the rat plasma and its application to a pharmacokinetic study

“…Speci city There was no interference with detection from blank plasma, since the plasma samples were subjected to 10-1000 folds dilution prior to analysis so as to greatly reduce interference from plasma matrix, as re ected by measured absorbance of blank plasma samples collected before administration of rLj-RGD3 being much lower than that of LOQ. More importantly, in our double sandwich BA-ELISA both capture antibody and detection antibody were mAbs, they combined with epitopes at two different sites, in this way, the speci city was greatly improved as compared with conventional ELISA where detection or capture antibody is usually a polyclonal antibody, as done for ELISA quanti cation of r-hirudin [8] , mCRP [9] , lactoferrin [10] , serum hepcidin [11] , tumor necrosis factor-α [12] , etc.…”

A picogram BA-ELISA quantification assay for rLj-RGD3, a platelet fibrinogen receptor antagonist, in the rat plasma and its application to a pharmacokinetic study

“…There was no interference in the detection process from blank plasma since the plasma samples were subjected to 10-1000 folds dilution prior to analysis so as to greatly reduce interference from the plasma matrix, as reflected by the much lower absorbance measured in the blank plasma samples collected before the administration of rLj-RGD3, compared to the limit of quantification. More importantly, in our double sandwich BA-ELISA, both the capture antibody and detection antibody were mAbs, they combined with epitopes at two different sites, in this way, the specificity was greatly improved as compared with conventional ELISA where detection or capture antibody is usually a polyclonal antibody, as done for ELISA PLOS NEGLECTED TROPICAL DISEASES quantification of r-hirudin [11], mCRP [12], lactoferrin [13], serum hepcidin [14], tumor necrosis factor-α [15], etc.…”

A picogram BA-ELISA quantification assay for rLj-RGD3, a platelet fibrinogen receptor antagonist, in the rat plasma and its application to a pharmacokinetic study

“…Speci city There was no interference with detection from blank plasma, since the plasma samples were subjected to 10-1000 folds dilution prior to analysis so as to greatly reduce interference from plasma matrix, as re ected by measured absorbance of blank plasma samples collected before administration of rLj-RGD3 being much lower than that of LOQ. More importantly, in our double sandwich BA-ELISA both capture antibody and detection antibody were mAbs, they combined with epitopes at two different sites, in this way, the speci city was greatly improved as compared with conventional ELISA where detection or capture antibody is usually a polyclonal antibody, as done for ELISA quanti cation of r-hirudin [8] , mCRP [9] , lactoferrin [10] , serum hepcidin [11] , tumor necrosis factor-α [12] , etc. Linearity and LOQ As illustrated in Fig.…”