Table Of ContentAptamers Engineered
Nanocarriers for Cancer
Therapy
Edited by
Prashant Kesharwani
Assistant Professor,
Department of Pharmaceutics,
SchoolofPharmaceuticalEducationandResearch,
Jamia Hamdard,
New Delhi, India
WoodheadPublishingisanimprintofElsevier
50HampshireStreet,5thFloor,Cambridge,MA02139,UnitedStates
TheBoulevard,LangfordLane,Kidlington,OX51GB,UnitedKingdom
Copyright©2023ElsevierLtd.Allrightsreserved.
Nopartofthispublicationmaybereproducedortransmittedinanyformorbyanymeans,
electronicormechanical,includingphotocopying,recording,oranyinformationstorage
andretrievalsystem,withoutpermissioninwritingfromthepublisher.Detailsonhowto
seekpermission,furtherinformationaboutthePublisher’spermissionspoliciesandour
arrangementswithorganizationssuchastheCopyrightClearanceCenterandthe
CopyrightLicensingAgency,canbefoundatourwebsite:www.elsevier.com/permissions.
Thisbookandtheindividualcontributionscontainedinitareprotectedundercopyrightby
thePublisher(otherthanasmaybenotedherein).
Notices
Knowledgeandbestpracticeinthisfieldareconstantlychanging.Asnewresearchand
experiencebroadenourunderstanding,changesinresearchmethods,professional
practices,ormedicaltreatmentmaybecomenecessary.
Practitionersandresearchersmustalwaysrelyontheirownexperienceandknowledgein
evaluatingandusinganyinformation,methods,compounds,orexperimentsdescribed
herein.Inusingsuchinformationormethodstheyshouldbemindfuloftheirownsafety
andthesafetyofothers,includingpartiesforwhomtheyhaveaprofessionalresponsibility.
Tothefullestextentofthelaw,neitherthePublishernortheauthors,contributors,or
editors,assumeanyliabilityforanyinjuryand/ordamagetopersonsorpropertyasamatter
ofproductsliability,negligenceorotherwise,orfromanyuseoroperationofanymethods,
products,instructions,orideascontainedinthematerialherein.
ISBN:978-0-323-85881-6
ForinformationonallWoodheadPublishingpublicationsvisitour
websiteathttps://www.elsevier.com/books-and-journals
Publisher:MatthewDeans
AcquisitionsEditor:SabrinaWebber
EditorialProjectManager:JoshuaMearns
ProductionProjectManager:PrasannaKalyanaraman
CoverDesigner:MilesHitchen
TypesetbyTNQTechnologies
Contributors
KhalilAbnous PharmaceuticalResearchCenter,PharmaceuticalTechnologyInsti-
tute,Mashhad University of MedicalSciences, Mashhad, Iran
MohammedA.S.Abourehab DepartmentofPharmaceutics,CollegeofPharmacy,
Umm Al-QuraUniversity,Makkah,SaudiArabia;DepartmentofPharmaceutics and
Industrial Pharmacy,College of Pharmacy,MiniaUniversity, Minia, Egypt
Barbara Adinolfi Istituto di Fisica Applicata Nello Carrara, Consiglio Nazionale
delleRicerche, Sesto Fiorentino, Italy
Sara Davari Ahranjani Department of Medical Nanotechnology, School of
Advanced Technologies in Medicine, Tehran University of Medical Sciences
(TUMS), Tehran, Iran
Abdulfattah Al-Kadash Cell Therapy Center, The University of Jordan, Amman,
Jordan
Bayan Abu Al-Ragheb Cell Therapy Center, The University of Jordan, Amman,
Jordan
MohammadSarwarAlam DepartmentofChemistry,SchoolofChemicalandLife
Sciences, Jamia Hamdard, New Delhi, Delhi,India
Walhan Alhaer CellTherapy Center, The University of Jordan,Amman, Jordan
MahnazAlipour PolymerCompositeResearchLaboratory,DepartmentofApplied
Chemistry,Faculty ofChemistry,University of Tabriz, Tabriz, Iran
Atul Anand Animal Disease Control Program, Department of Animal Husbandry
and Dairying, Ministry of Fisheries, Animal Husbandry & Dairying, Governemnt of
India, NewDelhi,India
PeymanAsadi DepartmentofMedicalNanotechnology,SchoolofAdvancedTech-
nologiesin Medicine, Mazandaran University ofMedical Sciences, Sari, Iran
MohammadBanazadeh PharmaceuticalSciencesandCosmeticProductsResearch
Center, Kerman University of Medical Sciences, Kerman, Iran; Students Research
Committee, Faculty of Pharmacy, Kerman University of Medical Sciences, Kerman,
Iran
xii Contributors
Mahmood Barani Medical Mycology and Bacteriology Research Center, Kerman
University ofMedical Sciences, Kerman, Iran
Jaleh Barar Research Center for Pharmaceutical Nanotechnology, Biomedicine
Institute,TabrizUniversityofMedicalSciences,Tabriz,EastAzerbaijan,Iran;Depart-
mentofPharmaceutics,FacultyofPharmacy,TabrizUniversityofMedicalSciences,
Tabriz, East Azerbaijan, Iran
PayamBayat FacultyofMedicine,DepartmentofImmunologyandAllergy,Immu-
nology Research Center, BuAli Research Institute, Mashhad University of Medical
Sciences, Mashhad, Iran
BehzadBehnam PharmaceuticsResearchCenter,InstituteofNeuropharmacology,
KermanUniversityofMedicalSciences,Kerman,Iran;FacultyofPharmacy,Students
ResearchCommittee,KermanUniversity ofMedical Sciences,Kerman,Iran; Herbal
andTraditionalMedicinesResearchCenter,KermanUniversityofMedicalSciences,
Kerman, Iran; Department of Pharmaceutical Biotechnology, Faculty of Pharmacy,
Kerman University ofMedical Sciences, Kerman, Iran
MagdolnaCasian DepartmentofAnalyticalChemistry,FacultyofPharmacy,Iuliu
Hațieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania
Cecilia Cristea Department of Analytical Chemistry, Faculty of Pharmacy, Iuliu
Hațieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania
Carla Cruz CICS-UBIdCentro de Investigaç~ao em Ciências da Sa(cid:2)ude, Universi-
dade da Beira Interior, Covilh~a, Portugal
Rambabu Dandela Department of Industrial and Engineering Chemistry, Institute
of Chemical Technology, Indian Oil Odisha Campus, Samantapuri, Bhubaneswar,
Odisha,India
SapnaDevi DivisionofVeterinaryMedicine,FacultyofVeterinaryScience&Ani-
malHusbandry,Sher-e-KashmirUniversityofAgriculturalSciences&Technologyof
Jammu(SKUAST-J),R.S. Pura,Jammu,India
Ezaldeen Esawi Cell Therapy Center, The University ofJordan, Amman, Jordan
MortezaEskandani ResearchCenterforPharmaceuticalNanotechnology,Biome-
dicineInstitute, TabrizUniversityofMedicalSciences,Tabriz,EastAzerbaijan,Iran
AidaGholoobi MedicalGeneticsResearchCenter,MashhadUniversityofMedical
Sciences,Mashhad,Iran;MetabolicSyndromeResearchCenter,MashhadUniversity
of Medical Sciences, Mashhad,Iran
Ambra Giannetti Istituto di Fisica Applicata Nello Carrara, Consiglio Nazionale
delle Ricerche,Sesto Fiorentino, Italy
LopamudraGiri DepartmentofIndustrialandEngineeringChemistry,Instituteof
Chemical Technology, Indian Oil Odisha Campus, Samantapuri, Bhubaneswar,Odi-
sha, India
Contributors xiii
ElaheGozali DepartmentofHealthInformationTechnology,SchoolofAlliedMed-
ical Sciences, Urmia University ofMedical Sciences, Urmia, Iran
Mukesh Kumar Gupta Department of Biotechnology and Medical Engineering,
National Instituteof Technology, Rourkela,Odisha,India
Farnaz Hosseini Department of Medical Nanotechnology, School of Advanced
Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz,
Iran
OanaHosu DepartmentofAnalyticalChemistry,FacultyofPharmacy,IuliuHație-
ganu University of Medicineand Pharmacy,Cluj-Napoca, Romania
Said I. Ismail Qatar Genome Project, Qatar Foundation, Doha, Qatar; Department
ofBiochemistryandPhysiology,MedicalSchool,TheUniversityofJordan,Amman,
Jordan
Ananya Kar Department of Industrial and Engineering Chemistry, Institute of
Chemical Technology, Indian Oil Odisha Campus, Samantapuri, Bhubaneswar, Odi-
sha, India
HarsimranKaur DepartmentofPharmaceutics,DelhiPharmaceuticalSciencesand
Research University, New Delhi,Delhi, India
GowthamKenguva DepartmentofIndustrialandEngineeringChemistry,Institute
of Chemical Technology, Indian Oil Odisha Campus, Samantapuri, Bhubaneswar,
Odisha, India
Prashant Kesharwani Department of Pharmaceutics, School of Pharmaceutical
Education and Research, Jamia Hamdard, New Delhi, Delhi, India; University Insti-
tute of Pharma Sciences, Chandigarh University, Mohali,Punjab, India
MehrdadKhatami NoncommunicableDiseases ResearchCenter, BamUniversity
ofMedicalSciences,Bam,Iran;FacultyofMedicalSciences,DepartmentofMedical
Biotechnology, Tarbiat Modares University, Tehran, Iran
ZainabLafi Facultyof Pharmacy, The Middle East University, Amman,Jordan
IsmailSamiMahmoud FacultyofAppliedMedicalSciences,DepartmentofMed-
ical Laboratory Sciences, The HashemiteUniversity, Zarqa,Jordan
MuhammedMajeed Sabinsa Corporation,EastWindsor,NJ, United States
Atena Mansouri Cellular and Molecular Research Center, Birjand University of
Medical Sciences, Birjand,Iran
André Miranda CICS-UBIdCentro de Investigaç~ao em Ciências da Sa(cid:2)ude, Uni-
versidade daBeira Interior, Covilh~a,Portugal
Mohammad Mohajeri Department of Pharmacology and Toxicology, School of
Pharmacy, Kerman University of Medical Sciences,Kerman, Iran
xiv Contributors
SeyedehAliaMoosavian NanotechnologyResearchCenter,PharmaceuticalTech-
nology Institute, Mashhad University of Medical Sciences, Mashhad, Razavi Khora-
san Province, Iran
Hamdi Nsairat Faculty of Pharmacy, Pharmacological and Diagnostic Research
Center, Al-Ahliyya Amman University, Amman, Jordan
Yadollah Omidi Department of Pharmaceutical Sciences, College of Pharmacy,
Nova Southeastern University, Fort Lauderdale, FL, United States
Hossein Omidian Department of Pharmaceutical Sciences, College of Pharmacy,
Nova Southeastern University, Fort Lauderdale, FL, United States
Kumar Pranay Department of Biochemistry, Indira Gandhi Institute of Medical
Sciences, Patna, Bihar, India
AlexandraPusta Department ofAnalytical Chemistry,FacultyofPharmacy,Iuliu
Hațieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania
Rajkumar Rajendram Department of Medicine, King Abdulaziz Medical City,
King Abdulaziz International Medical Research Center, Ministry of National Guard
eHealthAffairs,Riyadh,SaudiArabia;CollegeofMedicine,KingSaudBinAbdu-
laziz University ofHealthSciences, Riyadh, Saudi Arabia
SmrutiRekhaRout DepartmentofIndustrialandEngineeringChemistry,Institute
of Chemical Technology, Indian Oil Odisha Campus, Samantapuri, Bhubaneswar,
Odisha,India
Pratikshya Sa Laboratory of Nanomedicine, Institute of Life Sciences, Bhubanes-
war, Odisha, India; Regional Centre for Biotechnology, Faridabad, Haryana, India
Amirhossein Sahebkar Biotechnology Research Center, Pharmaceutical Technol-
ogy Institute, Mashhad University of Medical Sciences, Mashhad, Iran; Applied
Biomedical Research Center, Mashhad University of Medical Sciences, Mashhad,
Iran;SchoolofMedicine,TheUniversityofWesternAustralia,Perth,WA,Australia;
Department of Biotechnology, School of Pharmacy, Mashhad University of Medical
Sciences,Mashhad,Iran;FacultyofMedicine,DepartmentofMedicalBiotechnology
and Nanotechnology, Mashhad University of Medical Sciences,Mashhad, Iran
Sanjeeb Kumar Sahoo Laboratory of Nanomedicine, Institute of Life Sciences,
Bhubaneswar, Odisha, India
FatemehSalahpour-Anarjan DepartmentofMedicalNanotechnology,Facultyof
Advanced Medical Sciences, Tabriz University ofMedical Sciences, Tabriz, Iran
Kiarash Saleki Student Research Committee, Babol University of Medical Scien-
ces, Babol,Iran; USERNOffice,BabolUniversity of Medical Sciences, Babol, Iran
TiagoSantos CICS-UBIdCentrodeInvestigaç~aoemCiênciasdaSa(cid:2)ude,Universi-
dade da Beira Interior, Covilh~a, Portugal
Contributors xv
Thozhukat Sathyapalan Department of Academic Diabetes, Endocrinology and
Metabolism, Hull YorkMedical School, University ofHull, Hull, UnitedKingdom
Mahsa Shahriari Department of Pharmaceutical Biotechnology, Faculty of Phar-
macy,ZabolUniversityofMedicalSciences,Zabol,Iran;DepartmentofPharmaceut-
ical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences,
Mashhad, Iran; Pharmaceutical Research Center, Pharmaceutical Technology Insti-
tute,Mashhad University of MedicalSciences, Mashhad, Iran
NeeleshSharma DivisionofVeterinaryMedicine,FacultyofVeterinaryScience&
Animal Husbandry, Sher-e-Kashmir University of Agricultural Sciences & Technol-
ogy of Jammu (SKUAST-J), R.S. Pura, Jammu, India
AfsanaSheikh DepartmentofPharmaceutics,SchoolofPharmaceuticalEducation
and Research, Jamia Hamdard, New Delhi, Delhi, India
VanshikhaSingh DepartmentofPharmaceutics, School ofPharmaceuticalEduca-
tion and Research,Jamia Hamdard, New Delhi, India
Seyed Mohammad Taghdisi Targeted Drug Delivery Research Center, Pharma-
ceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad,
Iran
Yong Teng Department of Hematology and Medical Oncology, Winship Cancer
Institute, Emory University School of Medicine, Atlanta,GA, United States
Mihaela Tertis Department of Analytical Chemistry, Faculty of Pharmacy, Iuliu
Hațieganu University of Medicineand Pharmacy,Cluj-Napoca, Romania
SaraTombelli IstitutodiFisicaApplicataNelloCarrara,ConsiglioNazionaledelle
Ricerche, Sesto Fiorentino, Italy
Somayeh Vandghanooni Research Center for Pharmaceutical Nanotechnology,
BiomedicineInstitute,TabrizUniversityofMedicalSciences,Tabriz,EastAzerbaijan,
Iran
Rezvan Yazdian-Robati Molecular and Cell Biology Research Center, Faculty of
Medicine, Mazandaran University of Medical Sciences, Sari, Iran
FatemehYazdian FacultyofNewSciencesandTechnologies,DepartmentofLife
Science Engineering, University ofTehran,Tehran, Iran
FatemehZahedipour StudentResearchCommittee,FacultyofMedicine,Mashhad
University of Medical Sciences, Mashhad, Iran; Faculty of Medicine, Department of
MedicalBiotechnologyandNanotechnology,MashhadUniversityofMedicalScien-
ces, Mashhad, Iran
FarazZare Faculty of Medicine, Aalborg University, Aalborg, Denmark
1
Cell-SELEX technology for
aptamer selection
Gowtham Kenguva1, SmrutiRekha Rout1, LopamudraGiri1,
AmirhosseinSahebkar2,4,5, Prashant Kesharwani3,6 andRambabu Dandela1
1Department of Industrial and Engineering Chemistry, Institute of Chemical Technology,
IndianOil Odisha Campus, Samantapuri,Bhubaneswar, Odisha, India; 2Biotechnology
Research Center, Pharmaceutical TechnologyInstitute, Mashhad University of Medical
Sciences, Mashhad, Iran;3Department ofPharmaceutics, School of Pharmaceutical
Education and Research, Jamia Hamdard, New Delhi, Delhi, India; 4Applied Biomedical
ResearchCenter,MashhadUniversityofMedicalSciences,Mashhad,Iran;5Departmentof
Biotechnology, School of Pharmacy,MashhadUniversity of Medical Sciences, Mashhad,
Iran;6UniversityInstituteofPharmaSciences,ChandigarhUniversity,Mohali,Punjab,India
1.1 Introduction
Aptamers are oligonucleotides (short form of RNA or DNA) having multiple
applications in different fields like diagnostics, forensics, food science, research,
drugdiscovery,andgenetestingandarespeciallyusedfordrugdeliverycarrieragents
due to their special property affinity and target specificity [1]. The compounds with
highaffinityforthetargetmolecules,whicharefoundinahugecombinatorialnucleic
aciddatabase,areknownasAptamers.Theyareveryspecifictothetargetmoleculesas
wellastheyhavesomeotherqualitiessuchasrobustnesswhichopposesthereduction
conditionsaswellastheheat denaturation [2e6].Whencomparedtotheantibodies,
aptamers offer a different set of benefits, such as the preparation of chemicals is
easy, compact size, lower molecular weight, nonimmunogenicity, and simplicity
of alteration and ligation to nanoparticles for therapeutic, diagnosis, and imaging
applications [7].
Developing aptamers for the biomolecular target on the cell membrane in their
natural shape, like as proteins, molecular biomarkers, and receptors, has evolved as
a potential helpful technique to develop effective disease-specific probes [8]. A live
infectious organism, African trypanosomes also known as Trypanosoma brucei, was
usedinthefirstcell-SELEXstudy.Thefindingsrevealedthediscoveryofthreeclas-
sificationsofhighattractionRNAaptamerswhichareparticularfortheparasite’sin-
fectious bloodstream lifespan series phase. The aptamers are chosen to target the
parasite’sflagellarpocket,butnoneofthemattachtotheVSGprotein,thatisamong
themostcommonpolypeptidesontheTrypanosomainterface[9].Aptamershavebeen
shown in several investigations to not only attach to the cell external but also to be
carriedintracellularly.
AptamersEngineeredNanocarriersforCancerTherapy.https://doi.org/10.1016/B978-0-323-85881-6.00019-1
Copyright©2023ElsevierLtd.Allrightsreserved.
2 AptamersEngineeredNanocarriersforCancerTherapy
Aptamers are discovered using a continuous selectioneamplification process
knownasselectivegeneration ofligandsbyexponentialenrich.Intheprocess,culti-
vationoftargetmoleculewithasolostrandednucleicacidpoolwhichhave1014e1015
variations of randomized 30e100-nt sequenced takes place. After that, the variants
having required binding activity are redeemed, then the enriched library is amplified
by reverse transcriptionepolymerase chain reaction (PCR). The single-stranded pool
is then produced by deleting the template-strand, which can be done in-vitro as
well, where PCR product is generated. The whole procedure is repeated more than
20 times to identify aptamers [2]. The SELEX procedure has been established in
1930 that is selection of RNA molecules that bind to certain proteins in vitro [10].
RNA ligands to bacteriophage T4 DNA polymerase, systematic development of li-
gands through exponential enrichment, since then the various aptamers had been
created against different targets, which includes both small chemical molecules and
largemoleculeslikeproteins(multidomain).VariousSELEXprocesshasbeendevel-
oped by scientist which iscompatible with the requirement and makes the whole se-
lection procedure highly efficient. Capillary electrophoresis-SELEX, microfluidic
SELEX,rapid-SELEX,andcell-SELEXaresomeofthesemethods.Alloftheprevi-
ousSELEXapproaches,withtheexceptionofcell-SELEX,relyontargetinformation
for aptamer choice, and mostly aptamers found against pure protein targets fails to
identifythesameproteininitsnativeform.Toachievehighest aptamers, theprotein
must be kept in a homogeneous, naturally folded condition. SELEX has been done
directlyagainstpureplasmamembranesinordertochoiceaptamersundermorephys-
iologicalsettings[7].Intheyear1998,MorrisandJensenwerethefirsttousethecell-
basedSELEXapproachbywhichthemembranesofhumanredbloodcellswereused
as a complex mixture to identify aptamers that provides an in vitro procedure for
isolating high affinitive aptamers, especially againstacompound combinationofpo-
tential target compounds. Contrasting to the other SELEX approaches, cell-SELEX
choosesanaptamerincontradictionoftheentirecell,ensuringthatmoleculartargets
at cell membrane are in their natural condition and represent its normal folding
structure [8].
Incubation, partitioning, and amplification are the primary processes of
cell-SELEX,whicharecomparabletostandardSELEX(Fig.1.1).Positiveandnega-
tivecollectionsareusedinthecell-SELEXprocesstodevelopaptamersthatcanfight
cancer selectively. -ve selection is required to eliminate motifs that attach to normal
cellswhilealso improvingthe selectivity ofpotential aptamers.
Tobegin,withtheaptamer,asingle-strandedoligonucleotidecollectionwithawide
variety of randomly sequenced is generated and cultured with goal cells. Following
washing, the DNA fragments attached to the marked cell surface are extracted from
thecellsthroughheatingcell-ssDNAcomplexesathightemperaturesandcentrifuga-
tion.Andthentherestoredpoolandthenegativecontrolcellsarekeptfortheincuba-
tion and after that all the ssDNA which binds with the negative control cells are
separated. Utilizing a biotinylated reversing primer, unbinded motifs are enlarged
byPCR, resultingintheabsorption ofaparticularbinder tothetarget.Forcapturing
the separated biotinylated antisense strand as well as unlabeled ssDNA streptavidin
