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Rapid and Efficient Purification of Functional Collectin-12 and Its Opsonic Activity against Fungal Pathogens

Collectin-12 (collectin placenta 1, CL-P1, or CL-12) is a newly identified pattern recognition molecule of the innate immune system. Recent evidences show that CL-12 plays important roles not only in innate immune protection against certain clinically important pathogens but also in scavenging of ho...

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Autores principales: Zhang, Jie, Li, Anna, Yang, Chang-qing, Garred, Peter, Ma, Ying Jie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6701420/
https://www.ncbi.nlm.nih.gov/pubmed/31482100
http://dx.doi.org/10.1155/2019/9164202
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author Zhang, Jie
Li, Anna
Yang, Chang-qing
Garred, Peter
Ma, Ying Jie
author_facet Zhang, Jie
Li, Anna
Yang, Chang-qing
Garred, Peter
Ma, Ying Jie
author_sort Zhang, Jie
collection PubMed
description Collectin-12 (collectin placenta 1, CL-P1, or CL-12) is a newly identified pattern recognition molecule of the innate immune system. Recent evidences show that CL-12 plays important roles not only in innate immune protection against certain clinically important pathogens but also in scavenging of host molecules, leukocyte recruitment, and cancer metastasis. Furthermore, CL-12 has been shown to be associated with the pathogenesis of human diseases such as Alzheimer's disease and multiple sclerosis lesion development. Therefore, the functional consequence of CL-12 remains intriguing and awaits further elucidation. However, available protocols for the purification of recombinant CL-12 with high purity are laborious and inefficient and hamper further functional studies. Here, we report a simple, rapid, and efficient solution to obtain biologically active CL-12 with high purity. We established stable transfected Flp-In™-CHO cells expressing the recombinant CL-12 extracellular domain in high amounts. Recombinant CL-12 was purified from cell culture supernatants using a 3-step rapid purification procedure utilizing disposable affinity and ion exchange minicolumns. Purified recombinant CL-12 adopted an oligomeric structure with monomers, dimers, and trimers and retained its binding capacity towards the A. fumigatus strain that has been described before. Furthermore, we demonstrated the opsonic properties towards eight clinical isolates of A. fumigatus strains and diverse clinically important fungal pathogens. Purified recombinant CL-12 revealed a differential binding capacity towards selected fungal pathogens in vitro. In conclusion, we demonstrate a rapid and efficient purification solution for further biochemical and functional characterization of CL-12 and reveal opsonic properties of CL-12 towards diverse fungal pathogens.
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spelling pubmed-67014202019-09-03 Rapid and Efficient Purification of Functional Collectin-12 and Its Opsonic Activity against Fungal Pathogens Zhang, Jie Li, Anna Yang, Chang-qing Garred, Peter Ma, Ying Jie J Immunol Res Research Article Collectin-12 (collectin placenta 1, CL-P1, or CL-12) is a newly identified pattern recognition molecule of the innate immune system. Recent evidences show that CL-12 plays important roles not only in innate immune protection against certain clinically important pathogens but also in scavenging of host molecules, leukocyte recruitment, and cancer metastasis. Furthermore, CL-12 has been shown to be associated with the pathogenesis of human diseases such as Alzheimer's disease and multiple sclerosis lesion development. Therefore, the functional consequence of CL-12 remains intriguing and awaits further elucidation. However, available protocols for the purification of recombinant CL-12 with high purity are laborious and inefficient and hamper further functional studies. Here, we report a simple, rapid, and efficient solution to obtain biologically active CL-12 with high purity. We established stable transfected Flp-In™-CHO cells expressing the recombinant CL-12 extracellular domain in high amounts. Recombinant CL-12 was purified from cell culture supernatants using a 3-step rapid purification procedure utilizing disposable affinity and ion exchange minicolumns. Purified recombinant CL-12 adopted an oligomeric structure with monomers, dimers, and trimers and retained its binding capacity towards the A. fumigatus strain that has been described before. Furthermore, we demonstrated the opsonic properties towards eight clinical isolates of A. fumigatus strains and diverse clinically important fungal pathogens. Purified recombinant CL-12 revealed a differential binding capacity towards selected fungal pathogens in vitro. In conclusion, we demonstrate a rapid and efficient purification solution for further biochemical and functional characterization of CL-12 and reveal opsonic properties of CL-12 towards diverse fungal pathogens. Hindawi 2019-07-29 /pmc/articles/PMC6701420/ /pubmed/31482100 http://dx.doi.org/10.1155/2019/9164202 Text en Copyright © 2019 Jie Zhang et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Zhang, Jie
Li, Anna
Yang, Chang-qing
Garred, Peter
Ma, Ying Jie
Rapid and Efficient Purification of Functional Collectin-12 and Its Opsonic Activity against Fungal Pathogens
title Rapid and Efficient Purification of Functional Collectin-12 and Its Opsonic Activity against Fungal Pathogens
title_full Rapid and Efficient Purification of Functional Collectin-12 and Its Opsonic Activity against Fungal Pathogens
title_fullStr Rapid and Efficient Purification of Functional Collectin-12 and Its Opsonic Activity against Fungal Pathogens
title_full_unstemmed Rapid and Efficient Purification of Functional Collectin-12 and Its Opsonic Activity against Fungal Pathogens
title_short Rapid and Efficient Purification of Functional Collectin-12 and Its Opsonic Activity against Fungal Pathogens
title_sort rapid and efficient purification of functional collectin-12 and its opsonic activity against fungal pathogens
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6701420/
https://www.ncbi.nlm.nih.gov/pubmed/31482100
http://dx.doi.org/10.1155/2019/9164202
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