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Title of Journal: Appl Microbiol Biotechnol

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Abbravation: Applied Microbiology and Biotechnology

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Springer-Verlag

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DOI

10.1007/s00432-012-1203-5

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1432-0614

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High production of llama variable heavychain anti

Authors: Hiromoto Hisada Hiroko Tsutsumi Hiroki Ishida Yoji Hata
Publish Date: 2012/07/03
Volume: 97, Issue: 2, Pages: 761-766
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Abstract

Llama variable heavychain antibody fragment VHH fused to four different reader proteins was produced and secreted in culture medium by Aspergillus oryzae These fusion proteins consisted of Nterminal reader proteins VHH and a Cterminal histag sequence which facilitated purification using onestep histag affinity chromatography SDSPAGE analysis of the deglycosylated purified fusion proteins confirmed that the molecular weight of each corresponded to the expected sum of VHH and the respective reader proteins The apparent high molecular weight reader protein glucoamylase GlaB was found to be suitable for efficient VHH production The GlaBVHHHis protein bound its antigen human chorionic gonadotropin and was detectable by a new ELISAbased method using a coupled assay with glucoamylase glucose oxidase peroxidase maltose and 33′55′tetramethylbenzidine as substrates Addition of potassium phosphate to the culture medium induced secretion of 061 mg GlaBVHHHis protein/ml culture medium in 5 days

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