Mengke Yuan, Lan Yang, Chunyang Li, Zhiqiang Wu, Zhipeng Liu, Tianfei Du, Xinzong Rong, Cong Cong, Yongxia Zhang, Xiaoping Yu, Yali Gao, Zhengli Chen, Lanjun Liu, Yonghong Ge
{"title":"有效长效人α-Gal A-Fc融合蛋白在转基因小鼠乳汁中的表达。","authors":"Mengke Yuan, Lan Yang, Chunyang Li, Zhiqiang Wu, Zhipeng Liu, Tianfei Du, Xinzong Rong, Cong Cong, Yongxia Zhang, Xiaoping Yu, Yali Gao, Zhengli Chen, Lanjun Liu, Yonghong Ge","doi":"10.1007/s11248-025-00452-x","DOIUrl":null,"url":null,"abstract":"<p><p>Fabry disease is a rare X-linked inherited lysosomal storage disorder caused by a reduction or deficiency in the activity of α-galactosidase A (α-Gal A). The short half-life of α-Gal A necessitates biweekly infusions, thereby imposing significant economic and physical burdens on patients and their families. In this study, a novel long-acting replacement for α-Gal A, termed α-galactosidase A-Fc (α-Gal A-Fc), was designed. Two transgenic founders with an 18.2% transgene rate were obtained to express recombinant human α-Gal A-Fc protein in mouse milk. The α-Gal A-Fc enzyme activity in the milk of high-copy mice were significantly higher than those in low-copy mice and were stably inherited across F1-F3 generations. No significant differences were observed in α-Gal A-Fc concentration or enzymatic activity among high-copy mice of the same generation. During early lactation, the α-Gal A-Fc concentration and enzymatic activity were 2.1-fold and 2.17-fold higher, respectively, compared to late lactation. The expression levels during late lactation did not affect purification efficiency, allowing for the pooling of milk from high-copy mice throughout the entire lactation period for protein purification. The elimination half-life of the purified α-Gal A-Fc protein in mouse serum was 471 min, approximately 43 times longer than that of the commercially available drug Replagal. These findings facilitate the development of an efficient production system for long-acting human α-Gal A-Fc fusion protein and provide valuable insights into the utilization of transgenic large animal mammary gland bioreactors for biopharmaceuticals.</p>","PeriodicalId":23258,"journal":{"name":"Transgenic Research","volume":"34 1","pages":"33"},"PeriodicalIF":2.0000,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Competent expression of effective and long-acting human α-Gal A-Fc fusion protein in the milk of transgenic mice.\",\"authors\":\"Mengke Yuan, Lan Yang, Chunyang Li, Zhiqiang Wu, Zhipeng Liu, Tianfei Du, Xinzong Rong, Cong Cong, Yongxia Zhang, Xiaoping Yu, Yali Gao, Zhengli Chen, Lanjun Liu, Yonghong Ge\",\"doi\":\"10.1007/s11248-025-00452-x\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Fabry disease is a rare X-linked inherited lysosomal storage disorder caused by a reduction or deficiency in the activity of α-galactosidase A (α-Gal A). The short half-life of α-Gal A necessitates biweekly infusions, thereby imposing significant economic and physical burdens on patients and their families. In this study, a novel long-acting replacement for α-Gal A, termed α-galactosidase A-Fc (α-Gal A-Fc), was designed. Two transgenic founders with an 18.2% transgene rate were obtained to express recombinant human α-Gal A-Fc protein in mouse milk. The α-Gal A-Fc enzyme activity in the milk of high-copy mice were significantly higher than those in low-copy mice and were stably inherited across F1-F3 generations. No significant differences were observed in α-Gal A-Fc concentration or enzymatic activity among high-copy mice of the same generation. During early lactation, the α-Gal A-Fc concentration and enzymatic activity were 2.1-fold and 2.17-fold higher, respectively, compared to late lactation. The expression levels during late lactation did not affect purification efficiency, allowing for the pooling of milk from high-copy mice throughout the entire lactation period for protein purification. The elimination half-life of the purified α-Gal A-Fc protein in mouse serum was 471 min, approximately 43 times longer than that of the commercially available drug Replagal. These findings facilitate the development of an efficient production system for long-acting human α-Gal A-Fc fusion protein and provide valuable insights into the utilization of transgenic large animal mammary gland bioreactors for biopharmaceuticals.</p>\",\"PeriodicalId\":23258,\"journal\":{\"name\":\"Transgenic Research\",\"volume\":\"34 1\",\"pages\":\"33\"},\"PeriodicalIF\":2.0000,\"publicationDate\":\"2025-07-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Transgenic Research\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://doi.org/10.1007/s11248-025-00452-x\",\"RegionNum\":3,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Transgenic Research","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1007/s11248-025-00452-x","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
摘要
法布里病是一种罕见的x连锁遗传性溶酶体贮积症,由α-半乳糖苷酶a (α-Gal a)活性降低或缺乏引起。α-Gal A的半衰期短,需要每两周注射一次,这给患者及其家属带来了巨大的经济和身体负担。本研究设计了一种新的长效α-半乳糖苷酶α-半乳糖苷酶a - fc (α-Gal a - fc)。在小鼠乳中表达重组人α-Gal A-Fc蛋白,获得了两个转基因建立子,转基因率为18.2%。高拷贝小鼠乳中α-Gal A-Fc酶活性显著高于低拷贝小鼠,并在F1-F3代间稳定遗传。同一代高拷贝小鼠α-Gal A-Fc浓度和酶活性无显著差异。泌乳早期α-Gal A-Fc浓度和酶活性分别比泌乳后期高2.1倍和2.17倍。哺乳后期的表达水平不影响纯化效率,允许在整个哺乳期间汇集高拷贝小鼠的乳汁进行蛋白质纯化。纯化的α-Gal A-Fc蛋白在小鼠血清中的消除半衰期为471 min,比市售药物Replagal的消除半衰期长约43倍。这些发现促进了长效人α-Gal A-Fc融合蛋白高效生产体系的建立,并为转基因大型动物乳腺生物反应器在生物制药领域的应用提供了有价值的见解。
Competent expression of effective and long-acting human α-Gal A-Fc fusion protein in the milk of transgenic mice.
Fabry disease is a rare X-linked inherited lysosomal storage disorder caused by a reduction or deficiency in the activity of α-galactosidase A (α-Gal A). The short half-life of α-Gal A necessitates biweekly infusions, thereby imposing significant economic and physical burdens on patients and their families. In this study, a novel long-acting replacement for α-Gal A, termed α-galactosidase A-Fc (α-Gal A-Fc), was designed. Two transgenic founders with an 18.2% transgene rate were obtained to express recombinant human α-Gal A-Fc protein in mouse milk. The α-Gal A-Fc enzyme activity in the milk of high-copy mice were significantly higher than those in low-copy mice and were stably inherited across F1-F3 generations. No significant differences were observed in α-Gal A-Fc concentration or enzymatic activity among high-copy mice of the same generation. During early lactation, the α-Gal A-Fc concentration and enzymatic activity were 2.1-fold and 2.17-fold higher, respectively, compared to late lactation. The expression levels during late lactation did not affect purification efficiency, allowing for the pooling of milk from high-copy mice throughout the entire lactation period for protein purification. The elimination half-life of the purified α-Gal A-Fc protein in mouse serum was 471 min, approximately 43 times longer than that of the commercially available drug Replagal. These findings facilitate the development of an efficient production system for long-acting human α-Gal A-Fc fusion protein and provide valuable insights into the utilization of transgenic large animal mammary gland bioreactors for biopharmaceuticals.
期刊介绍:
Transgenic Research focusses on transgenic and genome edited higher organisms. Manuscripts emphasizing biotechnological applications are strongly encouraged. Intellectual property, ethical issues, societal impact and regulatory aspects also fall within the scope of the journal. Transgenic Research aims to bridge the gap between fundamental and applied science in molecular biology and biotechnology for the plant and animal academic and associated industry communities.
Transgenic Research publishes
-Original Papers
-Reviews:
Should critically summarize the current state-of-the-art of the subject in a dispassionate way. Authors are requested to contact a Board Member before submission. Reviews should not be descriptive; rather they should present the most up-to-date information on the subject in a dispassionate and critical way. Perspective Reviews which can address new or controversial aspects are encouraged.
-Brief Communications:
Should report significant developments in methodology and experimental transgenic higher organisms