C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
167/139, 167/45
C12Q 1/00 (2006.01) A61K 39/44 (2006.01) G01N 33/54 (1980.01)
Patent
CA 1172164
ABSTRACT OF THE DISCLOSURE A method for detecting the presence of antibiotics in milk which comprises the steps of: (a) contacting a solid matrix having attached thereto a purified immobilized antibody with a volume of milk and an enzyme-labeled antibiotic, said antibody being specific to said antihiotic: (b) separating the matrix from the milk and rinsing the matrix with water to remove excess milk and enzyme-labeled antibiotic (c) contacting the rinsed matrix with a substrate, said substrate in the presence of said enzyme-labeled antibiotic exhibiting a color change the amount of which is quantitatively related to the amount of enzyme- labeled antibiotic; and (d) measuring the amount of antibiotic present in said milk by comparing the color change in said substrate with a standard. Also provided is a method for producing purified antibodies for use in the foregoing detection method by: (a) covalently conjugating an antibiotic having a lactam-ring in the molecule to a protein capable of binding thereto through said lactam-ring; (b) injecting into a host animal capable of raising antibodies specific to said antibiotic the conjugate obtained in step (a) so as to raise said specific antibodies; (c) covalently conjugating the same antibiotic in step (a) to a second protein capable of binding thereto through said lactam-ring and different than the protein used in step (a) to form a second conjugate; (d) covalently binding said second conjugate to a solid matrix to form an affinity matrix for purifying the antibodies (e) isolating and purifying the specific antibodies raised in step (b) by contacting the host animal serum with the affinity matrix; and (f) recovering the specific antibodies in pure form.
374840
Brown Rodney J.
Swaisgood Harold E.
Research Triangle Institute
Ridout & Maybee Llp
LandOfFree
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