Detection of methionine- and alanine-recombinant bovine somatotropins and their induced antibodies in serum and milk of cows suggests blood-milk barrier specificity for these compounds

Smits, Nathalie G. E.; Blokland, Marco; Wubs, Klaas L.; Bovee, Toine F.H.; Albada, Bauke; Ginkel, L. A. van; Nielen, Michel W. F.

doi:10.3168/jds.2020-19209

Cited by 4 publications

(9 citation statements)

References 41 publications

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“…The antibody responses appear specific to rbGH and equivalent to serum responses [58,59]. A recent study reports the formation of rbST-induced antibodies in animals treated with either Ala-rbGH or Met-rbGH [37]. It was observed in both treatments that the rbGH-induced antibodies were transferred from blood to milk, showing no blood-milk barrier specificity for these antibodies.…”

Section: Indirect Analytical Strategiesmentioning

confidence: 94%

“…Extracting and purifying a specific protein like rbGH from such a complex environment requires an accurate knowledge of these components in order to define the appropriate physical-chemical conditions and parameters for a suitable and effective protocol. Based on the physico-chemical properties of rbGH described above (Mw, pI) and considering the very low levels of concentration expected in milk (1-2 mg/mL) [31,37], the main components that should be a problem for the efficient analysis of these residues in milk are detailed below.…”

Section: Specificity Of Determining Recombinant Bovine Growth Hormone Administration In Milk 21 Milk Compositionmentioning

confidence: 99%

“…If the strategies to detect rbGH administration in milk are the same as those applied in the blood, i.e., detection of indirect markers or detection of residues of the substance itself, the challenges are exacerbated. Indeed, the expected concentrations are at the ppb (µg/L) level [31,37], and milk is a biological matrix even more complex than blood. The following paragraphs provide an overview of the strategies developed so far and an assessment of their performance and applicability.…”

Section: Analytical Strategies In Milk To Detect Rbgh Administrationmentioning

confidence: 99%

“…In 2019, Welsh et al presented results of bGH measurements in incurred milk samples without, however, describing the protocol used and its performances [71]. More recently, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, involving a selective antibodies purification step, describes as performance a decision limit (CCα) of 2.3 ng/mL in milk [37]. This level of performance allowed the identification of Ala-rbGH in the milk of treated animals as early as day one after administration at a level of 11 mg/mL and to continue this detection above the CCα for six days.…”

Section: Direct Analytical Strategiesmentioning

confidence: 99%

“…This involves the use of an approach involving structural recognition phenomena as made possible by immuno-recognition. Such a strategy has already been efficiently applied in bovine serum [76] and recently described in milk [37]. Further, on-line tryptic digestion should be considered to reinforce robustness of this step, which suffers reproducibility issues and would also offer a higher throughput in the analysis.…”

Section: Prospects For Efficient Analysis Of Rbgh In Milkmentioning

confidence: 99%

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The Promise and Challenges of Determining Recombinant Bovine Growth Hormone in Milk

Raux

Bichon

Benedetto

et al. 2022

Foods

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Recombinant bovine growth hormone (rbGH) is produced in large quantities and widely used in a number of countries worldwide to stimulate milk production in dairy animals. The use of this compound in animal production is strictly regulated by food safety directives in force, in particular in the European Union (EU). Although analytical strategies for the detection of rbGH in blood have been successfully reported over the past 15 years, they do not fully answer the expectations of either competent authorities or industrials that would expect measuring its occurrence directly in the milk. As a matrix of excretion but also of consumption, milk appears indeed as the matrix of choice for detecting the use of rbGH in dairy animals. It also allows large volumes to be collected without presenting an invasive character for the animal. However, rbGH detection in milk presents several challenges, mainly related to the sensitivity required for its detection in a complex biological matrix. This review article presents the specific difficulties associated with milk and provides an overview of the analytical strategies reported in the literature and whether they concern indirect or direct approaches to the detection of rbGH administration to animals, with applications either for screening or confirmation purposes.

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Section: Indirect Analytical Strategiesmentioning

confidence: 94%

Section: Specificity Of Determining Recombinant Bovine Growth Hormone Administration In Milk 21 Milk Compositionmentioning

confidence: 99%

Section: Analytical Strategies In Milk To Detect Rbgh Administrationmentioning

confidence: 99%

Section: Direct Analytical Strategiesmentioning

confidence: 99%

Section: Prospects For Efficient Analysis Of Rbgh In Milkmentioning

confidence: 99%

See 3 more Smart Citations

The Promise and Challenges of Determining Recombinant Bovine Growth Hormone in Milk

Raux

Bichon

Benedetto

et al. 2022

Foods

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From targeted methods to metabolomics based strategies to screen for growth promoters misuse in horseracing and livestock: A review

Cloteau

Kaabia²,

Bizec

et al. 2023

Food Control

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Microsphere Peptide-Based Immunoassay for the Detection of Recombinant Bovine Somatotropin in Injection Preparations

et al. 2022

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The use of peptides in immunoassays can be favored over the use of the full protein when more cost effective or less toxic approaches are needed, or when access to the full protein is lacking. Due to restricted access to recombinant bovine somatotropin (rbST), a protein enhancing growth and lactating performances of livestock, which use has been banned in the EU, Canada and Australia (amongst others), we developed a peptide-based biorecognition assay on an imaging planar array analyzer. For this, we identified the rbST epitope that is responsible for binding to the rbST-targeting monoclonal antibody 4H12 (MAb 4H12) to be 115DLEEGILALMR125. This linear peptide was synthesized and coupled to microspheres, after which it was tested in a biorecognition competitive inhibition assay format. We observed IC50 values of approximately 0.11 μg mL−1, which are lower than observed for the full rbST protein (IC50 = 0.20 μg mL−1). Importantly, there was no binding with the scrambled peptide. Preliminary results of directly coupled peptides in a microsphere biorecognition assay for detection of rbST are presented. Real-life applicability for detection of somatotropins (STs) in injection preparations of bovine-, porcine- and equine ST are shown. This newly developed immunoassay strongly supports future developments of peptide-based immunoassays to circumvent the limited access to the full protein.

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Detection of methionine- and alanine-recombinant bovine somatotropins and their induced antibodies in serum and milk of cows suggests blood-milk barrier specificity for these compounds

Cited by 4 publications

References 41 publications

The Promise and Challenges of Determining Recombinant Bovine Growth Hormone in Milk

The Promise and Challenges of Determining Recombinant Bovine Growth Hormone in Milk

From targeted methods to metabolomics based strategies to screen for growth promoters misuse in horseracing and livestock: A review

Microsphere Peptide-Based Immunoassay for the Detection of Recombinant Bovine Somatotropin in Injection Preparations

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