{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,25]],"date-time":"2026-07-25T20:02:32Z","timestamp":1785009752119,"version":"3.55.0"},"reference-count":63,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2020,6,5]],"date-time":"2020-06-05T00:00:00Z","timestamp":1591315200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003093","name":"Ministry of Higher Education, Malaysia","doi-asserted-by":"publisher","award":["UPM\/700-2\/1\/TRGS\/5535704"],"award-info":[{"award-number":["UPM\/700-2\/1\/TRGS\/5535704"]}],"id":[{"id":"10.13039\/501100003093","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Two-dimensional electrophoretic (2DE)-based proteomics remains a powerful tool for allergenomic analysis of goat\u2019s milk but requires effective extraction of proteins to accurately profile the overall causative allergens. However, there are several current issues with goat\u2019s milk allergenomic analysis, and among these are the absence of established standardized extraction method for goat\u2019s milk proteomes and the complexity of goat\u2019s milk matrix that may hamper the efficacy of protein extraction. This study aimed to evaluate the efficacies of three different protein extraction methods, qualitatively and quantitatively, for the 2DE-proteomics, using milk from two commercial dairy goats in Malaysia, Saanen, and Jamnapari. Goat\u2019s milk samples from both breeds were extracted by using three different methods: a milk dilution in urea\/thiourea based buffer (Method A), a triphasic separation protocol in methanol\/chloroform solution (Method B), and a dilution in sulfite-based buffer (Method C). The efficacies of the extraction methods were assessed further by performing the protein concentration assay and 1D and 2D SDS-PAGE profiling, as well as identifying proteins by MALDI-TOF\/TOF MS\/MS. The results showed that method A recovered the highest amount of proteins (72.68% for Saanen and 71.25% for Jamnapari) and produced the highest number of protein spots (199 \u00b1 16.1 and 267 \u00b1 10.6 total spots for Saanen and Jamnapari, respectively) with superior gel resolution and minimal streaking. Six milk protein spots from both breeds were identified based on the positive peptide mass fingerprinting matches with ruminant milk proteins from public databases, using the Mascot software. These results attest to the fitness of the optimized protein extraction protocol, method A, for 2DE proteomic and future allergenomic analysis of the goat\u2019s milk.<\/jats:p>","DOI":"10.3390\/molecules25112625","type":"journal-article","created":{"date-parts":[[2020,6,9]],"date-time":"2020-06-09T06:34:16Z","timestamp":1591684456000},"page":"2625","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Optimization of Protein Extraction Method for 2DE Proteomics of Goat\u2019s Milk"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0423-8695","authenticated-orcid":false,"given":"Muzammeer","family":"Mansor","sequence":"first","affiliation":[{"name":"Laboratory of Food Safety and Food Integrity, Institute of Tropical Agriculture and Food Security, Universti Putra Malaysia, Serdang 43400 UPM, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5705-0223","authenticated-orcid":false,"given":"Jameel R.","family":"Al-Obaidi","sequence":"additional","affiliation":[{"name":"Department of Biology, Faculty of Science and Mathematics, Universiti Pendidikan Sultan Idris, Tanjong Malim 35900, Perak, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nurain Nadiah","family":"Jaafar","sequence":"additional","affiliation":[{"name":"Laboratory of Food Safety and Food Integrity, Institute of Tropical Agriculture and Food Security, Universti Putra Malaysia, Serdang 43400 UPM, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Intan Hakimah","family":"Ismail","sequence":"additional","affiliation":[{"name":"Department of Paediatrics, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang 43400 UPM, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Atiqah Farah","family":"Zakaria","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang 43400 UPM, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohd Azri Zainal","family":"Abidin","sequence":"additional","affiliation":[{"name":"Department of Paediatrics, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang 43400 UPM, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinap","family":"Selamat","sequence":"additional","affiliation":[{"name":"Laboratory of Food Safety and Food Integrity, Institute of Tropical Agriculture and Food Security, Universti Putra Malaysia, Serdang 43400 UPM, Selangor, Malaysia"},{"name":"Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, Serdang 43400 UPM, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Son","family":"Radu","sequence":"additional","affiliation":[{"name":"Laboratory of Food Safety and Food Integrity, Institute of Tropical Agriculture and Food Security, Universti Putra Malaysia, Serdang 43400 UPM, Selangor, Malaysia"},{"name":"Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, Serdang 43400 UPM, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9494-7291","authenticated-orcid":false,"given":"Nuzul Noorahya","family":"Jambari","sequence":"additional","affiliation":[{"name":"Laboratory of Food Safety and Food Integrity, Institute of Tropical Agriculture and Food Security, Universti Putra Malaysia, Serdang 43400 UPM, Selangor, Malaysia"},{"name":"Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, Serdang 43400 UPM, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,6,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.1177\/0009922816664512","article-title":"Cow\u2019s Milk Protein Allergy","volume":"55","author":"Mousan","year":"2016","journal-title":"Clin. 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