Fluorescent phycocyanin and fluorescent phycoerythrin and their application

A phycocyanin and phycoerythrin technology, applied in phycocyanin and phycoerythrin, telecommunications, medical, testing, scientific research, health care, cosmetic applications, food fields, can solve the specific description of non-fluorescent applications , There are no requirements and indicators for fluorescence emission, and no application fields are involved, so as to achieve the effects of excellent water solubility, enhanced immunity, and stable fluorescence.

Inactive Publication Date: 2003-05-14
刘维国
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Chinese patent application No. 94109295.X discloses a kind of "Arthrospira platensis phycocyanin and its application", without any requirements and indicators for fluorescence emission, and no specific description of fluorescence application
Chinese Patent Application No. 01115003.3 discloses an "Application of R-Phycoerythrin in the Preparation of Photosensitizers for Phototherapy Tumors", which only describes the use of R-Phycoerythrin in the preparation of photosensitizers, and does not involve Application field

Method used

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  • Fluorescent phycocyanin and fluorescent phycoerythrin and their application
  • Fluorescent phycocyanin and fluorescent phycoerythrin and their application
  • Fluorescent phycocyanin and fluorescent phycoerythrin and their application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1: the preparation method of fluorescent phycocyanin

[0020] Fresh algae—centrifugal drying—ultrasonic crushing—centrifugation to take supernatant—

[0021] Salting out—centrifugation to collect protein—dissolving with buffer—on ion exchange column, elution—

[0022] Molecular sieve chromatography—elution—collection of active peaks—freeze drying—

[0023] Obtain fluorescent phycocyanin finished product.

[0024] The fluorescent phycocyanin can emit strong fluorescence when excited by a suitable wavelength. It has a blue freeze-dried powder appearance, is easily soluble in water, has an isoelectric point of 4.8, and has a characteristic spectrum absorption peak of 620nm and a fluorescence emission peak of 650nm. see figure 1 , figure 1 is its spectral property. Its molecular structure is a carrier protein and a chromophore connected by a thioether bond --- a tetrapyrrole compound that extends linearly with an open chain, and its apoprotein monomer consis...

Embodiment 2

[0027] Example 2: Fluorescent phycoerythrin-labeled monoclonal antibody is used to detect hepatitis B virus surface antigen

[0028] 1. Fluorescent markers and antibodies

[0029] The fluorescent marker adopts fluorescent phycoerythrin, whose A 565 / A 280>5.0, good water solubility. The characteristic absorption peak is 565nm, the fluorescence excitation peak is 495nm, and the fluorescence emission peak is 578nm. The anti-HbsAg monoclonal antibodies are self-made in our laboratory, all of which are anti-a subtypes. This paired combination has been confirmed by the enzyme labeling method to have good sensitivity and high specificity.

[0030] 2. Marking method

[0031] The heterobifunctional reagent N-succinimide-3-(2-pyridyldithio)-propionate (SPDP) was used to pretreat fluorescent phycoerythrin and monoclonal antibody (Mab), respectively, After the mixed reaction, the cross-linked product-fluorescent antibody is obtained.

[0032] 3. Immune response

[0033] The solid-...

Embodiment 3

[0045] Example 3: Effect of phycocyanin on the growth of human blood cancer cell lines

[0046] The effects of Spirulina phycocyanin on the growth of human blood cancer cell lines HL-60, K-562 and U-937 were determined by semi-solid agar culture method and MTT assay. The three kinds of tumor cells were treated with different concentrations of Spirulina phycocyanin under in vitro culture conditions. The results showed that Spirulina phycocyanin had different degrees of inhibitory effects on these three tumor cells, and there was a concentration-dose effect. The effect is strong.

[0047] (1) It has a significant inhibitory effect on the growth of HL-60 cells at a higher concentration (100ul / ml);

[0048] (2) It has a significant inhibitory effect on the growth of K-562 cells at concentrations (100ul / ml and 30ug / ml);

[0049] (3) It has significant inhibitory effect on the growth of U-937 cells at the concentration (100ul / ml).

[0050] Anti-radiation effect: From the perspect...

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Abstract

The present invention discloses a fluorescent phycocyanin and a fluorescent phycaerythrin. The phycocyanin and the phycaerythrin have molecular structure connected to open-chained linear tetraphyrrolyl compound as carrier protein and chromophore via thioether bond, are extracted from algae and can emit strong fluorescence after being excitation of light with proper wavelength. The present invention also discloses serial uses of the phycocyanin and the phycaerythrin in fluorescent detection and immunological analysis, in preparing anticarcinogen medicine and food, in preparing medicine and food for resisting irradiation, strengthening immune, in preparing photon switch, and in preparing food and cosmetic additive.

Description

Technical field [0001] The invention belongs to algae protein and its application, more specifically, relates to a kind of phycocyanin and phycoerythrin extracted from algae that can emit strong fluorescence, and its application in scientific research, medical treatment, detection, telecommunications, health care, food, cosmetics applications in other fields. Background technique [0002] As early as the beginning of the last century, it was reported abroad that there were strongly fluorescent red, violet and blue proteins in cyanobacteria and red algae. In 1910, Kylin named these pigment proteins as "Phycochromo-proteins" for the first time. This light-harvesting pigment protein that appears in large quantities in red algae, blue-green algae and cryptophyta is the phycobiliproteins (Phycobiliproteins, PBP) that is currently in urgent need of development, which mainly includes phycoerythrin (Phycoerythrin, PE), phycocyanin (Phycocyanin, PC), phycoerythrocyanin (Phycoerythr...

Claims

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Application Information

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IPC IPC(8): A61K38/16A61P35/00C07K14/405G01N33/68H05B33/00
Inventor 龚兴国
Owner 刘维国
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