# US 2019/0376953 A1 · Method for cancer grading

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- Number: US 2019/0376953 A1
- Document ID: US20190376953A1
- Status: Published application
- Kind: A1
- Application number: 16/002,538
- Filed: 2018-06-07
- Published: 2019-12-12
- Earliest claimed: 2018-06-07
- Assignees: National Synchrotron Radiation Research Center
- Applicants: National Synchrotron Radiation Research Center
- Inventors: YAO-CHANG LEE; PEI-YU HUANG
- Web page: https://patentgrep.com/patent/US20190376953A1

## CPC

- G01N 33/5091: {for testing the pathological state of an organism}
- G01N 33/5759: {involving compounds localised on the membrane of tumour or cancer cells}
- G01N 33/5756: {involving tumour-associated glycolinkage [TAG]}
- G01N 2800/56: Staging of a disease; Further complications associated with the disease
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

The invention relates to a method for establishing an index for a given cancer grade. The method includes profiling glycan distribution pattern of a reference cancer cell sample; adsorbing the profiled reference cancer cell sample with adsorbents; measuring the amount of the adsorbents adhering onto the profiled reference cancer cell sample; and acquiring reference correlations between the glycan distribution pattern of the reference cancer cell sample and the amount of the adsorbents adhering onto the profiled reference cancer cell sample to form the index of the given cancer grade. An index for a given cancer grade and a method for grading cancer of a test cell sample are also provided.

## Claims

### Claim 1

A method for establishing an index of a given cancer grade, comprising:
providing a reference cancer cell sample, wherein the reference cancer cell sample has the given cancer grade and comprises glycan chains anchored on the surface of the reference cancer cell sample;
providing a first adsorbent and a second adsorbent, wherein the carbon number of the first adsorbent and the second adsorbent is from to 46, and the carbon number of the first adsorbent is less than that of the second adsorbent;
profiling glycan distribution pattern of the glycan chains anchored on the surface of the reference cancer cell sample;
adsorbing the profiled reference cancer cell sample with the first adsorbent and the second adsorbent, respectively;
measuring the amount of the first adsorbent adhering onto the profiled reference cancer cell sample and measuring the amount of the second adsorbent adhering onto the profiled reference cancer cell sample, respectively; and
acquiring a first reference correlation between the glycan distribution pattern of the reference cancer cell sample and the amount of the first adsorbent adhering onto the profiled reference cancer cell sample; acquiring a second reference correlation between the glycan distribution pattern of the reference cancer cell sample and the amount of the second adsorbent adhering onto the profiled reference cancer cell sample, wherein the first reference correlation and the second reference correlation form the index of the given cancer grade.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the cancer is colon cancer, breast cancer, gastric cancer, cervical cancer, prostate cancer, renal cancer, neuroendocrine tumor or oral cavity cancer.

### Claim 3 (depends on 1)

The method according to claim 1, wherein the first adsorbent or the second adsorbent is n-alkane or n-alkyl ester.

### Claim 4 (depends on 1)

The method according to claim 1, wherein the profiling, adsorbing and measuring steps comprise:
conducting a hydrolysis reaction for varied lengths of time to stepwise hydrolyze the glycan chains anchored on the reference cancer cell sample;
adsorbing the reference cancer cell sample hydrolyzed for varied time lengths with the first adsorbent and the second adsorbent, respectively;
measuring the amount of the first adsorbent and the second adsorbent adhering onto the reference cancer cell sample hydrolyzed for varied time lengths, respectively.

### Claim 5 (depends on 4)

The method according to claim 4, wherein the hydrolysis reaction is conducted with a hydrolysis reagent in the range of pH value from about −1 to about 6.5.

### Claim 6 (depends on 1)

The method according to claim 1, wherein the profiling, adsorbing and measuring steps comprise:
applying a series of bias voltages to the reference cancer cell sample;
adsorbing the reference cancer cell sample with applying the bias voltages with the first adsorbent and the second adsorbent, respectively, and adsorbing the reference cancer cell sample without applying the bias voltages with the first adsorbent and the second adsorbent, respectively;
measuring the amount of the first adsorbent adhering onto the reference cancer cell sample and the second adsorbent adhering onto the reference cancer cell sample with applying bias voltages, and measuring the amount of the first adsorbent adhering onto the reference cancer cell sample and the second adsorbent adhering onto the reference cancer cell sample without applying bias voltages, respectively.

### Claim 7 (depends on 6)

The method according to claim 6, wherein a series of bias voltages are from about 0 V to about 500 V.

### Claim 8 (depends on 1)

The method according to claim 1, which further comprises:
providing a normal cell sample, wherein the normal cell sample comprises intrinsic glycan chains anchored on the surface of the normal cell sample;
profiling glycan distribution pattern of the glycan chains anchored on the surface of the normal cell sample;
adsorbing the profiled normal cell sample with the first adsorbent and the second adsorbent, respectively;
measuring the amount of the first adsorbent adhering onto the profiled normal cell sample and measuring the amount of the second adsorbent adhering onto the profiled normal cell sample, respectively; and
acquiring a first control correlation between the glycan distribution pattern of the normal cell sample and the amount of the first adsorbent adhering onto the profiled normal cell sample; acquiring a second control correlation between the glycan distribution pattern of the normal cell sample and the amount of a second adsorbent adhering onto the profiled normal cell sample, respectively.

### Claim 9 (depends on 1)

An index for a given cancer grade, which is established by the method according to claim 1.

### Claim 10 (depends on 9)

A method for grading cancer of a test cell sample, comprising:
providing the test cell sample, wherein the test cell sample comprises glycan chains anchored on the surface of the test cell sample;
providing a first adsorbent and a second adsorbent, wherein the carbon number of the first adsorbent and the second adsorbent is from 20 to 46, and the carbon number of the first adsorbent is less than that of the second adsorbent;
profiling glycan distribution pattern of the glycan chains anchored on the surface of the test cell sample;
adsorbing the profiled test cell sample with the first adsorbent and the second adsorbent, respectively;
measuring the amount of the first adsorbent adhering onto the profiled test cell sample and measuring the amount of the second adsorbent adhering onto the profiled test cell sample, respectively;
acquiring a first test correlation between the glycan distribution pattern of the test cell sample and the amount of the first adsorbent adhering onto the profiled test cell sample; acquiring a second test correlation between the glycan distribution pattern of the test cell sample and the amount of the second adsorbent adhering onto the profiled test cell sample; and
determining a cancer grade of the test cell sample by comparing the first test correlation and the second test correlation with the index of the given cancer grade according to claim 9.

### Claim 11 (depends on 10)

The method according to claim 10, wherein the cancer is colon cancer, breast cancer, gastric cancer, cervical cancer, prostate cancer, renal cancer, neuroendocrine tumor or oral cavity cancer.

### Claim 12 (depends on 10)

The method according to claim 10, wherein the first adsorbent or the second adsorbent is n-alkane or n-alkyl ester.

### Claim 13 (depends on 10)

The method according to claim 10, wherein the profiling, adsorbing and measuring steps comprise:
conducting a hydrolysis reaction for varied lengths of time to stepwise hydrolyze the glycan chains anchored on the test cell sample;
adsorbing the test cell sample hydrolyzed for varied time lengths with the first adsorbent and the second adsorbent, respectively; and
measuring the amount of the first adsorbent and the second adsorbent adhering onto the test cell sample hydrolyzed for varied time lengths, respectively.

### Claim 14 (depends on 13)

The method according to claim 13, wherein the hydrolysis reaction is conducted with a hydrolysis reagent in the range of pH value from about −1 to about 6.5.

### Claim 15 (depends on 10)

The method according to claim 10, wherein the profiling, adsorbing and measuring steps comprise:
applying a series of bias voltages to the test cell sample;
adsorbing the test cell sample with applying the bias voltages with the first adsorbent and the second adsorbent, respectively, and adsorbing the test cell sample without applying the bias voltages with the first adsorbent and the second adsorbent, respectively;
measuring the amount of the first adsorbent adhering onto the test cell sample and the second adsorbent adhering onto the test cell sample with applying bias voltages, and measuring the amount of the first adsorbent adhering onto the test cell sample and the second adsorbent adhering onto the test cell sample without applying bias voltages, respectively.

### Claim 16 (depends on 15)

The method according to claim 15, wherein the series of bias voltages are from about 0 V to about 500 V.