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Microalbuminuria immunogen, preparation method of microalbuminuria immunogen, polyclonal antibody and preparation method and application of polyclonal antibody

A urine microalbumin and polyclonal antibody technology, applied in the field of biochemistry, can solve the problems of excess, missed detection, insufficient sensitivity and specificity, etc.

Active Publication Date: 2019-04-05
GUANGDONG FAPON BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, in clinical application, when mAlB high-value-positive samples are prepared by using mAlB polyclonal antibody as an immunogen to detect urine microalbumin in high-value positive samples, the high-value mAlB-positive samples The urine microalbumin in the urine exceeds the detection limit of the mAlB polyclonal antibody, and there is a missed detection
For mAlB low-value positive samples, the mAlB polyclonal antibody prepared by traditional urinary microalbumin as immunogen has insufficient sensitivity and specificity and is easily misjudged as positive.

Method used

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  • Microalbuminuria immunogen, preparation method of microalbuminuria immunogen, polyclonal antibody and preparation method and application of polyclonal antibody
  • Microalbuminuria immunogen, preparation method of microalbuminuria immunogen, polyclonal antibody and preparation method and application of polyclonal antibody
  • Microalbuminuria immunogen, preparation method of microalbuminuria immunogen, polyclonal antibody and preparation method and application of polyclonal antibody

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preparation example Construction

[0023] The preparation method of urinary microalbumin immunogen of one embodiment, comprises the following steps:

[0024] S110, purifying urinary microalbumin in urine.

[0025] Specifically, the step of purifying urinary microalbumin in urine includes the following operations:

[0026] S111, rough and pure.

[0027] Specifically, urine is subjected to salting out to obtain a crude product of urinary microalbumin.

[0028] In one of the embodiments, the urine is subjected to ammonium sulfate salting-out treatment to obtain crude urine microalbumin. Further, the saturation of ammonium sulfate is 40%-55%. Furthermore, the saturation of ammonium sulfate is 40%, 50% or 55%. Of course, in some other embodiments, other salts (such as sodium chloride, sodium sulfate, etc.) can also be used to perform salting-out treatment on the urine. It can be understood that, the method of obtaining the crude urinary microalbumin product can also be other methods well known in the art except...

Embodiment 1

[0066] urinary microalbumin in purified urine

[0067] (1) The urine from clinical patients in the department of nephrology was subjected to 50% saturated ammonium sulfate salting-out treatment to precipitate some miscellaneous proteins, and the supernatant was collected to obtain crude urine microalbumin.

[0068] (2) Dialyze the crude product of urinary microalbumin into 10mM phosphate buffered saline, centrifuge, take the supernatant and put it on an affic-blue affinity chromatography column (filler affic-blue comes from Bio-Rad, an American company), and pass through 0 %~20% B (phosphate buffered saline containing 1.4M NaCl, PBS) for linear elution, 100% B for one-step elution, collect the eluted fractions of affic-blue100% affinity chromatography, and then pass through a ProteinG column to collect the flow Wear the sample to obtain intermediate purified urine microalbumin.

[0069] (3) The intermediate purified urine microalbumin was subjected to ultrafiltration and conc...

Embodiment 2

[0072] Preparation of urinary microalbumin immunogen

[0073] (1) Adjust the concentration of urinary microalbumin prepared in Example 1 to 3 mg / mL, and then treat it in a hot water bath at 55°C for 30 minutes, then immediately treat it in an ice bath at 0°C under dark conditions for 2 hours to obtain urinary microalbumin immunogen.

[0074] (2) Test the reactivity of the urinary microalbumin immunogen obtained in (1) by turbidimetry. The test results of Example 2 are shown in Table 1.

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Abstract

The invention relates to microalbuminuria immunogen, a preparation method of the microalbuminuria immunogen, a polyclonal antibody, a preparation method of the polyclonal antibody, a detection reagentand test paper. The preparation method of the microalbuminuria immunogen comprises the following steps: microalbuminuria in urine is purified; and the microalbuminuria is subjected to heat treatmentand cold treatment to be partially denatured, and the microalbuminuria immunogen is obtained, wherein the heat treatment temperature is 55-65 DEG C, and the cold treatment temperature is 0-5 DEG C. The polyclonal antibody obtained through immunization of the microalbuminuria immunogen has high accuracy in microalbuminuria detecting.

Description

technical field [0001] The invention relates to the technical field of biochemistry, in particular to a urinary microalbumin immunogen, a preparation method thereof, a polyclonal antibody, a preparation method and application thereof. Background technique [0002] Urinary microalbumin (mAlB) is an important indicator for clinical evaluation of renal function damage, and biochemical detection based on a biochemical analyzer is an important means of detecting this indicator. Depending on the degree of disease of the subjects, the mAlB content in the clinical biochemical test results varies greatly. The mAlB content in the urine of normal people is about 0.01mg / mL, but the mAlB content in patients with severe nephropathy may be as high as 40mg / mL-50mg / mL. Therefore, in clinical application, when mAlB high-value-positive samples are prepared by using mAlB polyclonal antibody as an immunogen to detect urine microalbumin in high-value positive samples, the high-value mAlB-positiv...

Claims

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Application Information

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IPC IPC(8): C07K14/76C07K16/18G01N33/68
CPCC07K14/76C07K16/18G01N33/6827G01N2333/76
Inventor 柯拓
Owner GUANGDONG FAPON BIOTECH CO LTD
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