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Anti-egf like domain multiple 6 (EGFL6) antibodies and their applications in cancer diagnosis and treatment

One-EGFL6, antibody technology, applied in the field of anti-EGFL-like domain multiple 6 antibodies

Active Publication Date: 2020-10-16
TAIPEI MEDICAL UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, EGFL6 alone has not actually been used in clinical cancer diagnosis

Method used

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  • Anti-egf like domain multiple 6 (EGFL6) antibodies and their applications in cancer diagnosis and treatment
  • Anti-egf like domain multiple 6 (EGFL6) antibodies and their applications in cancer diagnosis and treatment
  • Anti-egf like domain multiple 6 (EGFL6) antibodies and their applications in cancer diagnosis and treatment

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0107] Example 1 Construction and biopanning of anti-EGFL6 single chain antibody library

[0108] Female white leghorn (Gallus domesticus) were immunized by intramuscular injection with 50 ug of recombinant EGF6 in an equal volume of Freund's complete adjuvant. Three additional immunizations were performed with incomplete adjuvant at 7 day intervals. After each immunization, chicken IgY antibodies in the yolk were collected and titrated by enzyme-linked immunosorbent assay (ELISA) to determine the presence of a humoral anti-EGFL6 antibody immune response. Egg yolk was isolated from nitrogen white using 10% dextran sulfate according to a published protocol (Akita, E.M. and Nakai, S. (1993). Production and purification of Fab'fragments from chicken egg yolk immunoglobulin Y (IgY). J Immunol Methods 162, 155-164) for IgY purification.

[0109]Based on previous reports (Andris-Widhopf, J., Rader, C., Steinberger, P., Fuller, R. and Barbas, C.F. 3rd (2000). Methods for the genera...

example 2

[0110] Binding analysis of example 2 anti-EGFL6 single chain antibody

[0111] E. coli expressed scFv antibodies were purified and incubated with EGFL6 protein immobilized on nitrocellulose membranes. Binding was then detected by the addition of goat anti-chicken IgY light chain followed by HRP-conjugated donkey anti-goat Ig antibody. After three washes, the membrane was developed using diaminobenzidine (DAB) substrate until the desired strength was achieved. figure 2 Recombinant full-length EGFL6 protein (lane EGFL6) is shown and analyzed by SDS-PAGE under reducing conditions using N-terminal truncation (lane EGFL6-F37) and C-terminal truncation (lane EGFL6-F396). figure 2 A and 2B are shown in western blots by E5scFv ( figure 2 A) and S12 scFv ( figure 2 B) The identified full-length EGFL6 protein and EGFL6-F396 truncated fragments.

example 3

[0112] EGFL6 expression in the serum of patient with colorectal cancer in example 3

[0113] Expression of epidermal growth factor-like protein 6 (EGFL6) was assessed in the sera of cancer patients.

[0114] Such as image 3 EGFL6 protein expression was measured by Western blot in lung, colon, ovarian, prostate, brain, uterine, and breast cancer cell lines as shown in . EGFL6 is not expressed in normal cell lines. These results indicate that EGFL6 can be found in cancer patients and that EGFL6 can be used as a cancer marker.

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PUM

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Abstract

Provided are an anti-EGF like domain multiple 6 antibody (anti-EGFL6 antibody) and cancer detection (or diagnosis) and treatment using the anti-EGFL6 antibody. Further provided are a single-chain antibody fragments (scFv) and humanized antibody, which have ability in binding to EGFL6 and in inhibiting angiogenesis and cancer cell growth.

Description

technical field [0001] The present invention is in the field of cancer detection (or diagnosis) and treatment. In particular, the present invention relates to anti-EGFL6 domain multiplex 6 antibodies (anti-EGFL6 antibodies) and cancer detection (or diagnosis) and treatment using the anti-EGFL6 antibodies. Background technique [0002] Cancer is the uncontrolled growth of abnormal cells anywhere in the body. Abnormal cells are called cancer cells, malignant cells, or tumor cells. Early diagnosis of cancer often increases the likelihood of successful treatment or cure of the disease. [0003] Biomarkers are potential tools for detecting cancer. EGFL6 is a member of the epidermal growth factor (EGF) repeat superfamily of proteins and consists of an EGF-like repeat region and an integrin association motif (RGD). EGFL6 promotes endothelial cell migration and angiogenesis. George Yeung et al. reported that EGFL6 encodes the expected signal peptide, indicating its secretion, a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395C07K16/28C07K16/30C07K16/22
CPCC07K16/005C07K16/22A61K2039/505C07K2317/24C07K2317/622A61P35/00A61K39/001131C07K2317/565
Inventor 李雨青潘秀玲皇甫维君黄翠琴魏柏立黄瀚立郑淳淳黄振侨蔡耿彰叶坤土
Owner TAIPEI MEDICAL UNIV
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