# US 2018/0024133 A1 · Methods for Detecting and for Treating Pancreatic Cancer > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2018/0024133 A1 - Document ID: US20180024133A1 - Status: Published application - Kind: A1 - Application number: 15/216,768 - Filed: 2016-07-22 - Published: 2018-01-25 - Earliest claimed: 2016-07-22 - Assignees: Van Andel Research Institute - Applicants: Van Andel Research Institute - Inventors: Brian B. Haab; Bryan Reatini - Web page: https://patentgrep.com/patent/US20180024133A1 ## CPC - G01N 33/57525: of the liver or pancreas - C12Y 204/01065: 3-Galactosyl-N-acetylglucosaminide 4-alpha-L-fucosyltransferase (2.4.1.65), i.e. alpha-1-3 fucosyltransferase - G01N 2333/4725: Mucins, e.g. human intestinal mucin - G01N 2333/91102: with definite EC number (2.4.1.-) - G01N 2440/38: addition of carbohydrates, e.g. glycosylation, glycation - 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) - C12Y: ENZYMES ## Abstract A method of diagnosing pancreatic cancer in a patient by detecting a level of one or more glycoforms of a Lewis antigen and a level of the one or more glycoforms of MUC5AC. The patient diagnosed with pancreatic cancer then may be treated for this disease. Also, a method for detecting a level of a glycan in a sample which includes using a capture reagent to immobilize the glycan on a substrate; exposing the immobilized glycan to a detection reagent; detecting the level of the immobilized glycan; and combining the biological sample with one or more pre-capture enzymes and/or exposing the immobilized glycan to one or more pre-detection enzymes. ## Claims ### Claim 1 A method of diagnosing pancreatic cancer in a human patient, said method comprising: obtaining a biological sample from the human patient, wherein the biological sample includes one or more glycoforms of a Lewis antigen and one or more glycoforms of a mucin 5AC (MUC5AC); detecting a level of the one or more glycoforms of the Lewis antigen and a level of the one or more glycoforms of MUC5AC in the biological sample; and diagnosing the patient with pancreatic cancer when the one or more glycoforms of the Lewis antigen and the one or more glycoforms of MUC5AC are at a different level than a statistically validated threshold for the one or more glycoforms of the Lewis antigen and the one or more glycoforms of MUC5AC. ### Claim 2 (depends on 1) The method of claim 1, wherein the one or more glycoforms of the Lewis antigen are sialyl-Lewis A:sialyl-Lewis A (sLeA:sLeA), sialyl-Lewis A:sulfated Lewis A (sLeA:sulfo-LeA), sialyl-Lewis A:sulfated sialyl-Lewis A (sLeA:sulfo-sLeA), sialyl-Lewis A:sialyl-Lewis X (sLeA:sLeX), sialyl-Lewis A:sulfated Lewis X (sLeA:sulfo-LeX), or sialyl-Lewis A:sulfated sialyl-Lewis X (sLeA:sulfo-sLeX). ### Claim 3 (depends on 1) The method of claim 1, wherein the one or more glycoforms of MUC5AC are MUC5AC:type 1 sialyl-N-acetyl-lactosamine (MUC5AC:sLacNAc t1), MUC5AC:type1type 2 sialyl-N-acetyl-lactosamine (MUC5AC:sLacNAc t1t2), MUC5AC:sialyl-Lewis A (MUC5AC:sLeA), MUC5AC:sialyl-Lewis X (MUC5AC:sLeX), MUC5AC:sulfated-Lewis A (MUC5AC:sulfo-LeA), MUC5AC:sulfated-Lewis X (MUC5AC:sulfo-LeX), MUC5AC:sulfated sialyl-Lewis A (MUC5AC:sulfo-sLeA), or MUC5AC:sulfated sialyl-Lewis X (MUC5AC:sulfo-sLeX). ### Claim 4 (depends on 2) The method of claim 2, wherein the one or more glycoforms of the Lewis antigen are sLeA:sulfo-sLeA and sLeA:sulfo-sLeX. ### Claim 5 (depends on 4) The method of claim 4, wherein the one or more glycoforms of MUC5AC are MUC5AC:sLeA, MUC5AC:sLacNAc t1 and MUC5AC:sulfo-sLeX. ### Claim 6 (depends on 4) The method of claim 4, wherein the one or more glycoforms of MUC5AC are MUC5AC:sLacNAc t1t2 and MUC5AC:sulfo-sLeX. ### Claim 7 (depends on 1) The method of claim 1, wherein the one or more glycoforms of the Lewis antigen and the one or more glycoforms of MUC5AC are detected with one or more glycan-binding proteins. ### Claim 8 (depends on 7) The method of claim 7, wherein the one or more glycoforms of the Lewis antigen is detected with an E-selectin lectin, and wherein the one or more glycoforms of MUC5AC is detected with a 7LE anti-Lewis antigen antibody, a TRA-1-60 anti-Lewis antigen antibody, or a Coprinopsis cinerea lectin 2 (CCL2). ### Claim 9 (depends on 1) The method of claim 1, wherein the level of the one or more glycoforms of the Lewis antigen and the level of the one or more glycoforms of MUC5AC in the biological sample are elevated as compared to the control levels of the one or more glycoforms of the Lewis antigen and the one or more glycoforms of MUC5AC. ### Claim 10 (depends on 1) The method of claim 1, wherein the sample is blood serum. ### Claim 11 (depends on 1) The method of claim 1, wherein the diagnosis differentiates pancreatic cancer from benign pancreatic disease. ### Claim 12 A method of diagnosing pancreatic cancer in a human patient, wherein the human patient has a pancreas, said method comprising: obtaining a biological sample from the human patient, wherein the biological sample includes one or more glycoforms of a Lewis antigen and one or more glycoforms of MUC5AC; detecting a level of the one or more glycoforms of the Lewis antigen and the one or more glycoforms of the MUC5AC in the biological sample; obtaining an image of the pancreas of the patient; and diagnosing the patient with pancreatic cancer when (a) the one or more glycoforms of the Lewis antigen and the one or more glycoforms of the MUC5AC are at a different level than a statistically validated threshold for the one or more glycoforms of the Lewis antigen and the one or more glycoforms of the MUC5AC and (b) the image of the pancreas indicates pancreatic cancer in the patient. ### Claim 13 (depends on 12) The method of claim 12, wherein the image is obtained by performing an ultrasound (US), computerized tomography (CT) scanning, or magnetic resonance imaging (MRI) of the pancreas of the patient. ### Claim 14 (depends on 12) The method of claim 12, wherein the one or more glycoforms of the Lewis antigen are sialyl-Lewis A:sialyl-Lewis A (sLeA:sLeA), sialyl-Lewis A:sulfated Lewis A (sLeA:sulfo-LeA), sialyl-Lewis A:sulfated sialyl-Lewis A (sLeA:sulfo-sLeA), sialyl-Lewis A:sialyl-Lewis X (sLeA:sLeX), sialyl-Lewis A:sulfated Lewis X (sLeA:sulfo-LeX), or sialyl-Lewis A:sulfated sialyl-Lewis X (sLeA:sulfo-sLeX). ### Claim 15 (depends on 12) The method of claim 12, wherein the one or more glycoforms of MUC5AC are MUC5AC:type 1 sialyl-N-acetyl-lactosamine (MUC5AC:sLacNAc t1), MUC5AC:type 1 type 2 sialyl-N-acetyl-lactosamine (MUC5AC:sLacNAc t1t2), MUC5AC:sialyl-Lewis A (MUC5AC:sLeA), MUC5AC:sialyl-Lewis X (MUC5AC:sLeX), MUC5AC:sulfated-Lewis A (MUC5AC:sulfo-LeA), MUC5AC:sulfated-Lewis X (MUC5AC:sulfo-LeX), MUC5AC:sulfated sialyl-Lewis A (MUC5AC:sulfo-sLeA), or MUC5AC:sulfated sialyl-Lewis X (MUC5AC:sulfo-sLeX). ### Claim 16 (depends on 12) The method of claim 12, wherein the diagnosis differentiates the pancreatic cancer from benign pancreatic disease. ### Claim 17 A method of diagnosing and treating pancreatic cancer in a subject, said method comprising: obtaining a biological sample from the human subject, wherein said biological sample includes one or more glycoforms of a Lewis antigen and one or more glycoforms of MUC5AC; detecting a level of the one or more glycoforms of the Lewis antigen and a level of the one or more glycoforms of the MUC5AC in the biological sample; diagnosing the subject with pancreatic cancer when the one or more glycoforms of the Lewis antigen and the one or more glycoforms of the MUC5AC in the biological sample are at a different level than a statistically validated threshold for the one or more glycoforms of the Lewis antigen and the one or more glycoforms of the MUC5AC; and administering a therapeutically effective amount of a treatment for pancreatic cancer to the diagnosed subject. ### Claim 18 (depends on 17) The method of claim 17, wherein the one or more glycoforms of the Lewis antigen are sialyl-Lewis A:sialyl-Lewis A (sLeA:sLeA), sialyl-Lewis A:sulfated Lewis A (sLeA:sulfo-LeA), sialyl-Lewis A:sulfated sialyl-Lewis A (sLeA:sulfo-sLeA), sialyl-Lewis A:sialyl-Lewis X (sLeA:sLeX), sialyl-Lewis A:sulfated Lewis X (sLeA:sulfo-LeX), or sialyl-Lewis A:sulfated sialyl-Lewis X (sLeA:sulfo-sLeX). ### Claim 19 (depends on 17) The method of claim 17, wherein the one or more glycoforms of MUC5AC are MUC5AC:type 1 sialyl-N-acetyl-lactosamine (MUC5AC:sLacNAc t1), MUC5AC:type1type 2 sialyl-N-acetyl-lactosamine (MUC5AC:sLacNAc t1t2), MUC5AC:sialyl-Lewis A (MUC5AC:sLeA), MUC5AC:sialyl-Lewis X (MUC5AC:sLeX), MUC5AC:sulfated-Lewis A (MUC5AC:sulfo-LeA), MUC5AC:sulfated-Lewis X (MUC5AC:sulfo-LeX), MUC5AC:sulfated sialyl-Lewis A (MUC5AC:sulfo-sLeA), or MUC5AC:sulfated sialyl-Lewis X (MUC5AC:sulfo-sLeX). ### Claim 20 (depends on 17) The method of claim 17, wherein the treatment is surgically resecting a pancreatic cystic lesion in the subject, applying radiation to the pancreatic cystic lesion, or administering a chemotherapeutic agent to the subject. ### Claim 21 A method of providing medical services for a human patient suspected of having or having pancreatic cancer, said method comprising: requesting a biological sample from and diagnostic information about the patient, wherein the diagnostic information is a level of one or more glycoforms of a Lewis antigen and a level of one or more glycoforms of MUC5AC; and administering a therapeutically effective amount of a treatment for pancreatic cancer when the diagnostic information indicates that the level of the one or more glycoforms of the Lewis antigen and the one or more glycoforms of MUC5AC in the biological sample are at a different level than a statistically validated threshold for the one or more glycoforms of the Lewis antigen and the one or more glycoforms of MUC5AC. ### Claim 22 (depends on 21) The method of claim 21, wherein the one or more glycoforms of the Lewis antigen are sialyl-Lewis A:sialyl-Lewis A (sLeA:sLeA), sialyl-Lewis A:sulfated Lewis A (sLeA:sulfo-LeA), sialyl-Lewis A:sulfated sialyl-Lewis A (sLeA:sulfo-sLeA), sialyl-Lewis A:sialyl-Lewis X (sLeA:sLeX), sialyl-Lewis A:sulfated Lewis X (sLeA:sulfo-LeX), or sialyl-Lewis A:sulfated sialyl-Lewis X (sLeA:sulfo-sLeX). ### Claim 23 (depends on 21) The method of claim 21, wherein the one or more glycoforms of MUC5AC are MUC5AC:type 1 sialyl-N-acetyl-lactosamine (MUC5AC:sLacNAc t1), MUC5AC:type1 type 2 sialyl-N-acetyl-lactosamine (MUC5AC:sLacNAc t1t2), MUC5AC:sialyl-Lewis A (MUC5AC:sLeA), MUC5AC:sialyl-Lewis X (MUC5AC:sLeX), MUC5AC:sulfated-Lewis A (MUC5AC:sulfo-LeA), MUC5AC:sulfated-Lewis X (MUC5AC:sulfo-LeX), MUC5AC:sulfated sialyl-Lewis A (MUC5AC:sulfo-sLeA), or MUC5AC:sulfated sialyl-Lewis X (MUC5AC:sulfo-sLeX). ### Claim 24 (depends on 21) The method of claim 21, wherein the treatment is surgically resecting a pancreatic cystic lesion in the subject, applying radiation to the pancreatic cystic lesion, or administering a chemotherapeutic agent to the subject. ### Claim 25 A method of monitoring treatment for pancreatic cancer in a human patient, comprising: requesting a first biological sample from and first diagnostic information about the patient, wherein the first diagnostic information is a level of one or more glycoforms of a Lewis antigen and a level of the one or more glycoforms of MUC5AC in the first biological sample; administering a therapeutically effective amount of a treatment for pancreatic cancer to the patient; after administering the therapeutically effective amount of the treatment for pancreatic cancer to the patient, requesting a second biological sample from and second diagnostic information about the patient, wherein the second diagnostic information is a level of the one or more glycoforms of the Lewis antigen and a level of the one or more glycoforms of MUC5AC in the second biological sample; and comparing the first diagnostic information and the second diagnostic information to determine whether the one or more glycoforms of the Lewis antigen and the one or more glycoforms of MUC5AC in the first biological sample are at a different level than the one or more glycoforms of the Lewis antigen and the one or more glycoforms of MUC5AC in the second biological sample. ### Claim 26 (depends on 25) The method of claim 25, wherein the one or more glycoforms of the Lewis antigen are sialyl-Lewis A:sialyl-Lewis A (sLeA:sLeA), sialyl-Lewis A:sulfated Lewis A (sLeA:sulfo-LeA), sialyl-Lewis A:sulfated sialyl-Lewis A (sLeA:sulfo-sLeA), sialyl-Lewis A:sialyl-Lewis X (sLeA:sLeX), sialyl-Lewis A:sulfated Lewis X (sLeA:sulfo-LeX), or sialyl-Lewis A:sulfated sialyl-Lewis X (sLeA:sulfo-sLeX); and wherein the one or more glycoforms of MUC5AC are MUC5AC:type 1 sialyl-N-acetyl-lactosamine (MUC5AC:sLacNAc t1), MUC5AC:type1type 2 sialyl-N-acetyl-lactosamine (MUC5AC:sLacNAc t1t2), MUC5AC:sialyl-Lewis A (MUC5AC:sLeA), MUC5AC:sialyl-Lewis X (MUC5AC:sLeX), MUC5AC:sulfated-Lewis A (MUC5AC:sulfo-LeA), MUC5AC:sulfated-Lewis X (MUC5AC:sulfo-LeX), MUC5AC:sulfated sialyl-Lewis A (MUC5AC:sulfo-sLeA), or MUC5AC:sulfated sialyl-Lewis X (MUC5AC:sulfo-sLeX). ### Claim 27 A method for detecting a level of a glycan in a subject, comprising: obtaining from the subject a biological sample having the glycan; providing a substrate having a capture reagent; exposing the biological sample to the capture reagent to immobilize the glycan in the biological sample on the substrate; providing a detection reagent; exposing the immobilized glycan to the detection reagent to bind the detection reagent to the immobilized glycan; visualizing the bound detection reagent to detect the level of the immobilized glycan; and performing one or both of following steps (a) and (b), wherein step (a) is, before exposing the biological sample to the capture reagent, combining one or more pre-capture enzymes with the biological sample, and wherein step (b) is, before exposing the immobilized glycan to the detection reagent, exposing the immobilized glycan to one or more pre-detection enzymes. ### Claim 28 (depends on 27) The method of claim 27, wherein the glycan is sialyl Lewis A. ### Claim 29 (depends on 28) The method of claim 28, wherein step (a) is performed and the one or more pre-capture enzymes is PNGaseF, or wherein step (b) is performed and the one or more pre-detection enzymes is PNGaseF, or wherein steps (a) and (b) both are performed and the one or more pre-capture enzymes is PNGaseF and the one or more pre-detection enzymes is PNGaseF. ### Claim 30 (depends on 27) The method of claim 27, wherein the glycan in the biological sample is indirectly immobilized on the substrate. ### Claim 31 (depends on 30) The method of claim 30, wherein the glycan is part of a glycoprotein or a glycolipid. ### Claim 32 (depends on 31) The method of claim 31, wherein the glycan is selected from the group consisting of sialyl Lewis A and sialyl N-acetyl-lactosamine type1. ### Claim 33 (depends on 32) The method of claim 32, wherein step (a) is performed and the capture reagent is an anti-Lewis A antibody and the one or more pre-capture enzymes is sialidase, or wherein step (b) is performed and the detection reagent is the anti-Lewis A antibody and the one or more pre-detection enzymes is sialidase, or wherein steps (a) and (b) both are performed and the capture reagent and the detection reagent both are the anti-Lewis A antibody and the one or more pre-capture enzymes and the one or more pre-detection enzymes both are sialidase. ### Claim 34 (depends on 33) The method of claim 33, wherein the anti-Lewis A antibody is a 7LE antibody. ### Claim 35 (depends on 34) The method of claim 34, wherein the glycan is part of a glycoprotein MUC5AC, the capture reagent is an anti-MUC5AC antibody, step (b) is performed and the detection reagent is the 7LE antibody, and the one or more pre-detection enzymes is sialidase. ### Claim 36 (depends on 31) The method of claim 31, wherein the glycan is sialyl N-acetyl-lactosamine type1. ### Claim 37 (depends on 36) The method of claim 36, wherein step (a) is performed and the capture reagent is an anti-sialyl N-acetyl-lactosamine type1 antibody and the one or more pre-capture enzymes is sialidase, or wherein step (b) is performed and the detection reagent is the anti-sialyl N-acetyl-lactosamine type1 antibody and the one or more pre-detection enzymes is sialidase, or wherein steps (a) and (b) both are performed and the capture reagent and the detection reagent both are the anti-sialyl N-acetyl-lactosamine type1 antibody and the one or more pre-capture enzymes and the one or more pre-detection enzymes both are sialidase. ### Claim 38 (depends on 37) The method of claim 37, wherein the anti-sialyl N-acetyl-lactosamine type1 antibody is a TRA-1-60 antibody or a TRA-1-81 antibody. ### Claim 39 (depends on 38) The method of claim 38, wherein the glycan is part of a glycoprotein MUC5AC, the capture reagent is an anti-MUC5AC antibody, step (b) is performed and the detection reagent is the TRA-1-60 antibody, and the one or more pre-detection enzymes is sialidase. ### Claim 40 (depends on 27) The method of claim 27, wherein the pre-capture enzyme is sialydase, PNGaseF, or a sulfatase. ### Claim 41 (depends on 27) The method of claim 27, wherein the pre-detection enzyme is sialydase, PNGaseF, or a sulfatase. ### Claim 42 (depends on 27) The method of claim 27, wherein the biological sample is blood or serum. ### Claim 43 (depends on 27) The method of claim 27, wherein the substrate is a microarray slide. ### Claim 44 (depends on 27) The method of claim 27, wherein the capture reagent is a glycan binding protein, or wherein the detection reagent is a glycan binding protein, or wherein the capture reagent and the detection reagent both are a glycan binding protein. ### Claim 45 (depends on 27) The method of claim 27, further comprising, after the biological sample is exposed to the capture reagent, washing the substrate to remove any extraneous material that is not immobilized on the substrate. ### Claim 46 (depends on 27) The method of claim 27, wherein the exposing the biological sample to the capture reagent step includes incubating the biological sample on the substrate. ### Claim 47 (depends on 27) The method of claim 27, wherein the exposing the immobilized glycan to the detection reagent step includes incubating the detection reagent on the substrate. ### Claim 48 (depends on 27) The method of claim 27, wherein step (b) is performed, and wherein the exposing the immobilized glycan to the one or more pre-detection enzymes includes incubating the one or more pre-detection enzymes on the substrate. ### Claim 49 (depends on 27) The method of claim 27, wherein the level of the immobilized glycan that is detected is different than a level of the immobilized glycan that is detected both in the absence of the combining the one or more pre-capture enzymes with the biological sample and in the absence of exposing the immobilized glycan to the one or more pre-detection enzymes. ### Claim 50 (depends on 49) The method of claim 49, wherein the level of the immobilized glycan that is detected is increased as compared to a level of the immobilized glycan that is detected both in the absence of the combining the one or more pre-capture enzymes with the biological sample and in the absence of exposing the immobilized glycan to the one or more pre-detection enzymes.