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Myoglobin immunoadsorbent and immunoadsorption device

An immunoadsorbent and myoglobin technology, which is applied in the field of immunoadsorption devices, can solve the problems of inability to achieve specific removal of myoglobin and affect blood components, and achieve the effects of easy implementation, low water and electricity consumption, and low degree of dependence

Inactive Publication Date: 2012-07-04
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are few literatures on the adsorption and removal of myoglobin. Kuntsevich et al. conducted a myoglobin adsorption test on the commercialized adsorber X-Sorb and found that the adsorption rate is good, but the adsorber is not specially designed for myoglobin and cannot achieve myoglobin adsorption. Specific clearance of hemoglobin, may affect other blood components
This study proposes a myoglobin-specific removal method using antibodies as ligands to construct an immunosorbent device, but there is no literature report yet

Method used

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  • Myoglobin immunoadsorbent and immunoadsorption device
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  • Myoglobin immunoadsorbent and immunoadsorption device

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1, the preparation of myoglobin polyclonal antibody

[0049] 1. Design and synthesis of short antigenic peptides

[0050] 1. Obtain the full-length amino acid sequence information of myoglobin

[0051] Search the full-length amino acid sequence of myoglobin in the NCBI protein database, which is shown in SEQ ID: No.1:

[0052] MGLSD GEWQM VLNIW GKVEG DLAGH GQEVL ISLFK AHPET LEKFD KFKNL KSEEE MKS SE DLKKH GCTVL TALGT ILKKK GQHAA EIQPL AQSHA TKHKI PVKYL EFISE VIIQV LKKRY SGDFG ADAQG AMSKA LELFR NDIAA KYKEL GFQG

[0053] 2. Epitope prediction

[0054] Refer to the bl2seq protein homology detection results on the NCBI website (http: / / blast.ncbi.nlm.nih.gov), and use DNAStar7.10 software to assist in the design of polypeptide antigens. Select antigenic peptides according to the following principles:

[0055] ①Located on the surface of the protein as much as possible; ②Good hydrophilicity; ③N-terminal or C-terminal is better than the middle segment; ④Generally,...

Embodiment 2

[0145] Example 2. Construction and functional verification of myoglobin immunoadsorption experimental treatment device

[0146] 1. Assemble the myoglobin immunoadsorption column

[0147] 1. Selection of carrier

[0148] Select the resin carrier according to the following basic conditions:

[0149] ①Material with -NH 2 ②neutral or weak acid-base; ③has a certain strength; ④low swelling rate; ⑤does not release biotoxic substances.

[0150] In this embodiment, weakly basic anion exchange resin D-380 is selected as the resin carrier. Serum incubation test was carried out on D-380 resin, and it was found that immersion in D-380 resin had no significant effect on the concentration of several main electrolytes in serum, such as Figure 4 As shown, it can be used as a carrier.

[0151] 2. Antibody Conjugation

[0152] The purified myoglobin polyclonal antibody was used as ligand, coupled with D-380 resin carrier to prepare adsorption column packing, the specific steps are as foll...

Embodiment 3

[0217] Embodiment 3, practical application of myoglobin immunoadsorption experimental treatment device

[0218] One, the first aid kit containing the immunoadsorption device of the present invention

[0219] 1. The immunosorbent device of the present invention can be prepared together with some necessary accessories into a first aid kit (Rescue Kit), which is used for on-site first aid for patients with excessive myoglobin (such as being crushed in accidents such as earthquakes and landslides). Generally, the first aid kit should include:

[0220] ① Myoglobin adsorption column; ② Large venous catheter; ③ Vascular puncture needle; ④ Gloves;

[0221] 2. The main steps when the first aid kit is applied to the human body are:

[0222] (1) Use catheter to establish vascular access;

[0223] (2) The extracted venous blood is pumped into the adsorption device through the blood pump, or the arterial blood is drawn directly into the adsorption device;

[0224] (3) After being absor...

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Abstract

The invention discloses a myoglobin immunoadsorbent which comprises a solid phase carrier and a myoglobin polyclonal antibody coupled with the carrier; the myoglobin polyclonal antibody is a polyclonal antibody obtained by using a compound as an immunogen, wherein the compound is formed by crosslinking keyhole limpet hemocyanin with antigenic peptide; the amino acid sequence of the antigenic peptide is shown in sequence table SEQ ID:No.5. The invention also discloses a myoglobin immunoadsorption column which uses the myoglobin immunoadsorbent as a filler, a myoglobin immunoadsorption device containing the myoglobin immunoadsorption column, and a first aid tool kit. The immunoadsorbent and the device of the invention are established in a convenient blood perfusion treatment mode, can realize myoglobin adsorption with high specificity, and are especially suitable for on-site rescue of rhabdomyolysis. Compared with modes of hemodialysis or hemofiltration, the whole blood perfusion mode has less water and electricity consumption, is easier to realize, and more meets the requirements of disaster accident on-site rescue.

Description

technical field [0001] The invention relates to an immunoadsorbent and an immunoadsorbent device containing the adsorbent, in particular to an immunoadsorbent device applicable to on-site first aid for rhabdomyolysis. Background technique [0002] Rhabdomyolysis can lead to acute kidney injury, and myoglobin is the key protein of pathogenesis, which can lead to the apoptosis of renal tubular epithelial cells. Clearing blood myoglobin is an effective way to treat rhabdomyolysis. [0003] Most of the hydrophobic side chains of the myoglobin polypeptide chain are inside the molecule, forming a bag-shaped cavity, and the hydrophilic side chains are mostly located on the surface of the molecule, so it has good water solubility and can be freely filtered by the glomerulus. When it is abnormally high, it is usually treated with hemodialysis or hemofiltration. However, traditional hemodialysis is not enough to remove myoglobin, and hemofiltration, especially high-flux hemofiltrati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395C07K19/00C07K17/08A61M1/38A61F17/00A61P7/08A61P21/00
Inventor 周建辉陈香美张利孙雪峰
Owner GENERAL HOSPITAL OF PLA
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