Multilectin assay for detecting fibrosis-specific glyco-alteration by means of lectin microarray.
BACKGROUND
Despite the progress made in understanding glyco-alterations of specific glycoproteins such as α1-acid glycoprotein (AGP) associated with liver fibrosis, there has been no useful diagnostic assay with a lectin recognizing the fibrosis-specific alteration and an antibody against the core protein. We therefore developed a compatible multiple lectin-antibody sandwich immunoassay on the basis of the results obtained by the lectin microarray analysis for monitoring fibrosis.


METHODS
AGP-enriched fractions derived from 0.5-μL sera of 125 patients with staging-determined fibrosis (26.4% F0-F1, 25.6% F2, 24% F3, and 23.2% F4) were subjected to systematic analysis by antibody-overlay lectin microarray. Data were analyzed to statistically relate to the degree of fibrosis progression. Additionally, we applied an optimal lectin signal set on the microarray to distinguish 45 patients with cirrhosis from 43 patients with chronic hepatitis.


RESULTS
Signal patterns of the 12 selected lectins reflected fibrosis-associated glyco-alteration of AGP. Among the 12 lectins, we found a specific lectin at each stage of fibrosis (i.e., significant fibrosis, severe fibrosis, and cirrhosis) (P < 0.0001). The test for the detection of cirrhosis showed that combinational use of 3 lectins (AOL, MAL, and DSA) on the array enhanced the diagnostic value for liver cirrhosis to 95% diagnostic sensitivity and 91% diagnostic specificity.


CONCLUSIONS
The multiple lectin-antibody sandwich immunoassay targeting AGP enables monitoring of disease progression in chronic hepatitis patients at risk of developing hepatocellular carcinoma.
Research Center for Medical Glycoscience (RCMG), National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, Japan.
Atsushi
A
Kuno
Yuzuru
Y
Ikehara
Yasuhito
Y
Tanaka
Takashi
T
Angata
Sachiko
S
Unno
Maki
M
Sogabe
Hidenori
H
Ozaki
Kiyoaki
K
Ito
Jun
J
Hirabayashi
Masashi
M
Mizokami
Hisashi
H
Narimatsu
D000328:Adult; D000911:Antibodies, Monoclonal; D018450:Disease Progression; D005260:Female; D006521:Hepatitis, Chronic / Q000097:blood; D006801:Humans; D007118:Immunoassay; D037102:Lectins; D008103:Liver Cirrhosis / Q000097:blood / Q000175:diagnosis* / Q000503:physiopathology; D008297:Male; D046228:Microarray Analysis / Q000379:methods; D008875:Middle Aged; D009961:Orosomucoid / Q000032:analysis* / Q000276:immunology; D011134:Polysaccharides / Q000097:blood; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D037102:Lectins; D009961:Orosomucoid; D011134:Polysaccharides