[Development and application of a method for rapid quantification of recombinant ICOS-mIg fusion protein].
OBJECTIVE
To set up a double antibody sandwich ELISA for rapid quantification of recombinant ICOS-mIg fusion protein and evaluate its sensitivity, specificity and linear detection range.


METHODS
Recombinant ICOS-mIg fusion protein was prepared by molecular biology technique. Sheep anti-mouse IgG and horse anti-mouse IgG-HRP conjugate were selected as coating antibody and detection antibody, respectively for sandwich ELISA by pair-matching experiments. A standard curve for determining the recombinant ICOS-mIg fusion protein was plotted and applied for quantification of ICOS-mIg fusion protein.


RESULTS
A sandwich ELISA for quantification of the recombinant ICOS-mIg fusion protein was developed. Its linear detection was 7.8 to 500 microg/L(R(2)=0.9911, P<0.0001).


CONCLUSIONS
The recombinant ICOS-mIg fusion protein can be determined by this sandwich ELISA rapidly and precisely.
Wang
Jian
J
Central Laboratory of Changhai Hospital, Second Military Medical University, Shanghai 200433, China.
Shen
Qian
Q
D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte / Q000235:genetics*; D015153:Blotting, Western; D016466:CHO Cells; D054370:Chemokine CXCL9 / Q000235:genetics*; D006224:Cricetinae; D003412:Cricetulus; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D060889:Inducible T-Cell Co-Stimulator Protein; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000945:Antigens, Differentiation, T-Lymphocyte; D054370:Chemokine CXCL9; C556692:ICOS protein, human; C556694:Icos protein, mouse; D060889:Inducible T-Cell Co-Stimulator Protein; D011993:Recombinant Fusion Proteins