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Violacein cải thiện sản xuất IgG tái tổ hợp bằng cách kiểm soát chu kỳ tế bào của tế bào buồng trứng hamster Trung Quốc
Tóm tắt
Tế bào buồng trứng hamster Trung Quốc (CHO) được sử dụng làm tế bào chủ cho sản xuất kháng thể đơn dòng (mAb) công nghiệp. Kiểm soát chu kỳ tế bào là một phương pháp hiệu quả để tăng cường sản xuất mAb trong nuôi cấy tế bào. Violacein, một sắc tố màu tím được sản xuất bởi vi sinh vật, có nhiều đặc tính sinh học hoạt động và đã được đề xuất cho nhiều ứng dụng công nghiệp khác nhau. Trong nghiên cứu này, chúng tôi đã đánh giá tiềm năng của violacein trong việc kiểm soát chu kỳ tế bào và cải thiện sản xuất immunoglobulin G (IgG) tái tổ hợp trong tế bào CHO. So với nhóm đối chứng, 0.9 μM violacein trong nuôi cấy theo phương pháp cho ăn 14 ngày đã làm tăng nồng độ IgG tối đa lên 37.6% thông qua việc tăng tỷ lệ sản xuất cụ thể và tuổi thọ tế bào. Phân tích chu kỳ tế bào cho thấy violacein đã gây ra tình trạng ngừng G1 và G2/M. Tuy nhiên, tình trạng ngừng G1 chỉ được quan sát vào ngày thứ nhất, trong khi tình trạng ngừng G2/M kéo dài hơn 3 ngày, gợi ý rằng tình trạng ngừng G2/M đã làm trung gian việc tăng cường sản xuất IgG do violacein gây ra. Hơn nữa, song song với việc tăng cường biểu hiện protein, mức độ biểu hiện mRNA của IgG và tỷ lệ chuyển hóa dinh dưỡng cũng đã được cải thiện. Glycosyl hóa liên kết N và hồ sơ biến thể điện tích hầu như không bị ảnh hưởng bởi điều trị bằng violacein. Kết quả của chúng tôi cho thấy violacein ảnh hưởng đến chu kỳ tế bào của tế bào CHO và tăng cường sản xuất IgG mà không làm thay đổi chất lượng sản phẩm, cho thấy tiềm năng như một tác nhân tăng cường sản xuất mAb trong tế bào CHO. Nghiên cứu này cung cấp cái nhìn sâu sắc về việc sử dụng violacein trong sản xuất mAb công nghiệp và có thể giúp phát triển các công nghệ sản xuất mAb tiên tiến và hiệu quả hơn bằng cách sử dụng nuôi cấy tế bào CHO.
Từ khóa
#violacein #tế bào CHO #mAb #IgG #kiểm soát chu kỳ tế bàoTài liệu tham khảo
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