Abstract
Immunoglobulin in yolk (IgY) (with a titer of 1.3 × 106) specific against bovine milk (BM) alkaline phosphatase (ALP) was obtained by intramuscularly immunizing hens on the thigh and was used as the primary antibody to conduct competitive indirect enzyme-linked immunosorbent assay (CI-ELISA) to determine BMALP in ALPs from BM and Escherichia coli sources. A relationship between the ELISA value and the BMALP level (0.01-10 μg/mL) in whole milk (R2 = 0.9019) or in skimmed milk (R2 = 0.9402) was observed. The maximal inhibition (%) of BMALP on the microtiter plate by free BMALP at 10 μg/mL whole milk (3.89 mU/μg BMALP) was about 50%, while no inhibition (%) of BMALP by free E. coli ALP at concentrations between 0.01 to 10 μg/mL (60 mU/μg E. coli ALP) was determined. At BMALP levels higher than 0.1 μg/mL, CI-ELISA was proved to be effective in differentiating between BMALP and E. coli ALP and quantifying BMALP in whole milk or skimmed milk in the presence of E. coli ALP with an activity of 0.6 U/mL. Higher inhibition (about 70%) of BMALP on the microtiter plate by free BMALP in diluted (101-104 fold) milk samples was observed. The optimal conditions for CI-ELISA in determining BMALP (0.1-10 μg/mL) from ALPs in milk samples were using 103-fold diluted crude IgY specific against BMALP as primary antibody and 103-fold diluted goat anti-chicken IgG-ALP conjugate as the secondary antibody.
| Original language | English |
|---|---|
| Pages (from-to) | 213-220 |
| Number of pages | 8 |
| Journal | Food Chemistry |
| Volume | 95 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 2006 Mar 1 |
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Keywords
- Alkaline phosphatase
- Bovine milk
- Competitive ELISA
- Escherichia coli
- Immunoglobulin in yolk
ASJC Scopus subject areas
- Analytical Chemistry
- Food Science
Cite this
Detection of alkaline phosphatase by competitive indirect ELISA using immunoglobulin in yolk (IgY) specific against bovine milk alkaline phosphatase. / Chen, Chao Cheng; Tai, Yu Chang; Shen, Szu-Chuan; Tu, Yann Ying; Wu, Ming Chang; Chang, Hung Min.
In: Food Chemistry, Vol. 95, No. 2, 01.03.2006, p. 213-220.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Detection of alkaline phosphatase by competitive indirect ELISA using immunoglobulin in yolk (IgY) specific against bovine milk alkaline phosphatase
AU - Chen, Chao Cheng
AU - Tai, Yu Chang
AU - Shen, Szu-Chuan
AU - Tu, Yann Ying
AU - Wu, Ming Chang
AU - Chang, Hung Min
PY - 2006/3/1
Y1 - 2006/3/1
N2 - Immunoglobulin in yolk (IgY) (with a titer of 1.3 × 106) specific against bovine milk (BM) alkaline phosphatase (ALP) was obtained by intramuscularly immunizing hens on the thigh and was used as the primary antibody to conduct competitive indirect enzyme-linked immunosorbent assay (CI-ELISA) to determine BMALP in ALPs from BM and Escherichia coli sources. A relationship between the ELISA value and the BMALP level (0.01-10 μg/mL) in whole milk (R2 = 0.9019) or in skimmed milk (R2 = 0.9402) was observed. The maximal inhibition (%) of BMALP on the microtiter plate by free BMALP at 10 μg/mL whole milk (3.89 mU/μg BMALP) was about 50%, while no inhibition (%) of BMALP by free E. coli ALP at concentrations between 0.01 to 10 μg/mL (60 mU/μg E. coli ALP) was determined. At BMALP levels higher than 0.1 μg/mL, CI-ELISA was proved to be effective in differentiating between BMALP and E. coli ALP and quantifying BMALP in whole milk or skimmed milk in the presence of E. coli ALP with an activity of 0.6 U/mL. Higher inhibition (about 70%) of BMALP on the microtiter plate by free BMALP in diluted (101-104 fold) milk samples was observed. The optimal conditions for CI-ELISA in determining BMALP (0.1-10 μg/mL) from ALPs in milk samples were using 103-fold diluted crude IgY specific against BMALP as primary antibody and 103-fold diluted goat anti-chicken IgG-ALP conjugate as the secondary antibody.
AB - Immunoglobulin in yolk (IgY) (with a titer of 1.3 × 106) specific against bovine milk (BM) alkaline phosphatase (ALP) was obtained by intramuscularly immunizing hens on the thigh and was used as the primary antibody to conduct competitive indirect enzyme-linked immunosorbent assay (CI-ELISA) to determine BMALP in ALPs from BM and Escherichia coli sources. A relationship between the ELISA value and the BMALP level (0.01-10 μg/mL) in whole milk (R2 = 0.9019) or in skimmed milk (R2 = 0.9402) was observed. The maximal inhibition (%) of BMALP on the microtiter plate by free BMALP at 10 μg/mL whole milk (3.89 mU/μg BMALP) was about 50%, while no inhibition (%) of BMALP by free E. coli ALP at concentrations between 0.01 to 10 μg/mL (60 mU/μg E. coli ALP) was determined. At BMALP levels higher than 0.1 μg/mL, CI-ELISA was proved to be effective in differentiating between BMALP and E. coli ALP and quantifying BMALP in whole milk or skimmed milk in the presence of E. coli ALP with an activity of 0.6 U/mL. Higher inhibition (about 70%) of BMALP on the microtiter plate by free BMALP in diluted (101-104 fold) milk samples was observed. The optimal conditions for CI-ELISA in determining BMALP (0.1-10 μg/mL) from ALPs in milk samples were using 103-fold diluted crude IgY specific against BMALP as primary antibody and 103-fold diluted goat anti-chicken IgG-ALP conjugate as the secondary antibody.
KW - Alkaline phosphatase
KW - Bovine milk
KW - Competitive ELISA
KW - Escherichia coli
KW - Immunoglobulin in yolk
UR - http://www.scopus.com/inward/record.url?scp=25144462448&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=25144462448&partnerID=8YFLogxK
U2 - 10.1016/j.foodchem.2005.01.028
DO - 10.1016/j.foodchem.2005.01.028
M3 - Article
AN - SCOPUS:25144462448
VL - 95
SP - 213
EP - 220
JO - Food Chemistry
JF - Food Chemistry
SN - 0308-8146
IS - 2
ER -