High-efficiency production of human serum albumin in the posterior silk glands of transgenic silkworms, Bombyx mori L.

Qiujie Qian,Yiran Wang,Shaohua Wang,Boxiong Zhong,Jiaqian Che,Zhengying You,Lupeng Ye,J Joe Hull

doi:10.1371/journal.pone.0191507

Abstract

Human serum albumin (HSA) is an important biological preparation with a variety of biological functions in clinical applications. In this study, the mRNA of a fusion transposase derived from the pESNT-PBase plasmid and a pBHSA plasmid containing the HSA gene under the control of a fibroin light chain (FL) promoter were co-injected into fertilized eggs. Fifty-six transgenic silkworm pedigrees expressing theexogenous recombinant HSA (rHSA) in the posterior silk glands (PSGs) with stable inheritance were successfully obtained. The SDS-PAGE and Western blot results confirmed that the rHSA was secreted into the transgenic silkworm cocoon, and the rHSA could be easily extracted with phosphate-buffered saline (PBS). In our research, the isolated highest amount rHSA constituted up to 29.1% of the total soluble protein of the cocoon shell, indicating that the transgenic silkworm produced an average of 17.4 μg/mg of rHSA in the cocoon shell. The production of soluble rHSA in the PSGs by means of generating transgenic silkworms is a novel approach, whereby a large amount of virus-free and functional HSA can be produced through the simple rearing of silkworms.

Highlights

Human serum albumin (HSA) is a non-glycosylated globulin, accounting for more than 50% of plasma proteins, and is the main factor contributing to blood colloid osmotic pressure
The SDSPAGE and Western blot results confirmed that the recombinant HSA (rHSA) was secreted into the transgenic silkworm cocoon, and the rHSA could be extracted with phosphate-buffered saline (PBS)
The isolated highest amount rHSA constituted up to 29.1% of the total soluble protein of the cocoon shell, indicating that the transgenic silkworm produced an average of 17.4 μg/mg of rHSA in the cocoon shell

Summary

Introduction

HSA is a non-glycosylated globulin, accounting for more than 50% of plasma proteins, and is the main factor contributing to blood colloid osmotic pressure. HSA can be used to transport drugs and metabolic factors because of its multiple binding sites [1,2], and is an important biological product in modern medicine. It is widely used in blood volume expansion and human protein supplementation, especially in the treatment of shock due to blood loss, trauma and burns, in mitigating cerebral hypertension caused by brain edema and brain damage, and in preventing hypoalbuminemia, cirrhosis, and kidney disease [3].

Methods

Results

Conclusion