Immune-colloidal gold test strip for detecting adulterant cow milk in camel milk and application thereof
A technology of colloidal gold test paper and camel milk, which is applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of non-existence, expensive instrument operation, and high sensitivity
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Embodiment 1
[0060] Example 1: SDS-PAGE compares the protein composition of cow milk and camel milk
[0061] Camel milk whey, bovine milk whey, camel milk casein, and bovine milk casein were subjected to polyacrylamide gel electrophoresis (SDS-PAGE), Coomassie brilliant blue stained gel and decolorized to record the results with a gel imager, see attached figure 1 As shown, it was found that camel whey lacked β-lactoglobulin relative to bovine whey, and the protein content of α-lactalbumin in camel whey was significantly higher than that in bovine whey. Therefore, bovine milk β-lactoglobulin and camel milk α-lactalbumin were selected for Preparation of polyclonal antibody; Camel milk casein contains less κ-casein and α-casein than bovine milk s1 - Casein has a large molecular weight, so cow's milk κ casein and camel milk α are selected s1 - Casein for the preparation of polyclonal or monoclonal antibodies.
Embodiment 2
[0062] Example 2: Camel milk α-lactalbumin and α s1 Extraction of casein
[0063] (1) Crude extraction of camel milk α-lactalbumin by ammonium sulfate fractional precipitation
[0064] Take defatted camel milk, adjust the pH to 4.3-4.8, centrifuge at 3000-12000r / min for 10-60min, and repeat the centrifugation several times. The supernatant is subjected to fractional precipitation according to the ammonium sulfate saturation of 70%-100%, and the obtained precipitate is desalted and freeze-dried spare.
[0065] (2) Calcium ion precipitation crude extraction of camel milk α s1 -casein
[0066] Take defatted camel milk, adjust the pH to 10-12, add calcium ions to produce a precipitate, adjust the pH to 6-8, centrifuge at 3000-8000r / min for 10-30min, take the precipitate and dissolve it in 3.3mol / L urea, adjust the pH to 4 ~5, centrifuge at 3000~8000r / min for 10~30min, get the precipitate and dry it for later use.
[0067] (3) Ultrafiltration and centrifugation for coarse sepa...
Embodiment 3
[0071] Example 3: Cow milk β-lactoglobulin, cow milk κ-casein, camel milk α-lactalbumin, camel milk α s1 - Preparation of monoclonal antibody against casein
[0072] (1) Bovine milk β-lactoglobulin, bovine milk κ-casein, camel milk α-lactalbumin, camel milk α s1 - Preparation of monoclonal antibody against casein:
[0073] Female Balb / c mice about 7 weeks old were selected as immunized animals, and the immunogens were bovine milk β-lactoglobulin, bovine milk κ-casein, camel milk α-lactalbumin and camel milk α s1 - Casein, immunogen and Freund's complete adjuvant (primary immunization) or Freund's incomplete adjuvant (boost immunization) immunization, after ultrasonic mixing and emulsification, multi-point subcutaneous and intraperitoneal injection to immunize mice. The titer of the isolated serum was determined by indirect ELISA method. When the titer of the serum reached above 1:64000, the mouse with the highest reaction value was injected into the tail vein and peritoneal ...
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