Torea Technologies                              

PO Box 823 Dunedin New Zealand

torea@netaccess.co.nz                                             

TOREA TECHNOLOGIES IS ENGAGED IN RESEARCH, DESIGN AND DEVELOPMENT OF  BIOTECHNOLOGY DERIVED ANTIBODY PHARMACEUTICALS

PRODUCTS ARE CURRENTLY DEVELOPED FOR CANCER CARE APPLICATIONS

 

CYTOTOXICS ADC

 

IMAGING RADIOPHARMACEUTICALS

                          MRI CONTRAST AGENTS 

                          RADIOTHERAPY

 

BIOACTIVE IMMUNOCONJUGATES WITH IMPROVED TARGETING PROPERTIES ARE ABLE TO BE UTILIZED IN CHEMOTHERAPY,RADIOTHERAPY,MRI AND NUCLEAR MEDICINE IMAGING

 


 
     
BIOACTIVE
IMMUNOCONJUGATE
PHARMACEUTICALS

CLINICAL APPLICATIONS  IN ONCOLOGY-CANCER THERAPY AND IMAGING
-IMPROVEMENTS IN TARGETING OF
BIOTECHNOLOGY DERIVED MONOCLONAL ANTIBODY BASED AGENTS
 

BIOSELECTIVITY CHARACTERISTICS

PURITY, POTENCY, IMMUNOCOGNIZANCE,  BINDING, AFFINITY, PERSISTENCE

Delivery augmentation by bioselective immunoconjugates of cytotoxics,mri and nuclear medicine agents
Higher tumor uptake,reduction of side effects and adverse delivery

BIOSELECTIVE IMMUNOREACTIVITY IMMUNOCONJUGATE
DESIGN PREPARATION

-High specific modification whilst remaining immunoreactive. Binding site protection allows for application of conjugation chemistry. Bound antibody is able to withstand greater concentrations of reactants for higher specific modification.Conjugation applied to the antibody in final bound conformation.
-Bioprocess preparation to allow folding competency.Folding changes upon binding accomodated with this product.
-Preparitive affinity purification with epitope based solid phase immunoadsorbent column bioprocess for production.

PRODUCTION OF A BIOACTIVE ANTIBODY AGENT RESULTING IN TARGETING WITH HIGH SUBSTITUENT MODIFICATION FOLDING CAPAPACITY AND FUNCTIONAL BINDING SITE RECOGNITION. PRODUCTION FOR PIPELINE DEVELOPMENT IN CHEMOTHERAPY,IMAGING AND ANTIBIOTICS. USEFUL IN TREATMENT RESISTANCE AND SOURCED FROM BIOTECHNOLGY AND MOLECULAR BIOLOGY RESEARCH.
 

CONTEMPORARY ANTIBODY CONJUGATION LIMITATIONS

Poor targeting,reduced immunoreactivity,low potency,low solubility,damaged folding properties....impurity loss of specificity and of limited clinical value.

The antibody is relatively inert chemically requiring high stoichiometric reactant concentrations with subsequent damage for conjugation.The folded state of free native antibody is metastable to ensure solubility,presentation of the binding site and binding affinity. Folding changes occur with binding as demonstrated with thermodynamic determination and X-ray crystallography studies.

 

Clinical limitations 1.Low targeting 2.Reduced binding 3.Low potencytocy 4.High background 5.Corrupted biodistribution
 

CONVENTIONAL ANTIBODY CONJUGATES EG BEXXAR AND ZEVALIN SUFFER FROM THE LOW IMMUNOREACTIVE FRACTION LOCALISING AND THE HIGH BACKGROUND LEADING TO COMPROMISED IMAGE RESOLUTION NEEDED FOR EARLY CANCER DETECTION. IN THERAPY THIS CAN MEAN INEFFECTIVE TREATMENT AND ADVERSE TOXICITY. IMPROVEMENTS IN EFFICACY CAN BE ACHIEVED WITH THESE AGENTS WITH BIOACTIVE LABELING TECHNOLOGY AND BIOPROCESS.

NATURE OF LABELING DAMAGE-IMPURITY

Labeling of the sensitive binding site where intimate contact is required.

Reactant chemical damage

Folding distortion particularly tertiary structure

Low retention and persistence upon binding to antigenic target

Low specific activity

Low solubility-particularly when higher substitution is attempted.

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MARKERS OF INTEREST

Cancer Pathologies....antigens and epitopes    

Lung....calcitonin,bombesin,cea etc

Prostate....psa, p-specific membrane antigen

Others....Ovary,melanoma,breast,leukaemia,colorectal....

Non cancer pathologies... Hiv....components of replication....

infection....bacterial viral
 

Epitope analysis required for refined specificity.

Cancer markers offer a therapeutic window via antigen density and relative specificity.

Epitope mapping of antigens has refined specificity eg TNF epitopes

Growth factor receptors eg EGFR,Cell surface antigens eg glycoproteins and secretory products such as hormones eg calcitonin,oncofoetal antigens eg CEA.

Availability from biotechnology sources ...... culture,fermentation,peptide,carbohydrate and glycosynthesis.

 

ANTIBODY AGENTS OF INTEREST

SOURCE-Biotechnology,fermentation,culture,monoclonals

FORMS-fragments,single chain,recombinant derivatives....phage libraries 

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CONJUGATION BIOSELECTIVITY PROCESS

 

A SYNTHETIC PATHWAY OF REACTIONS AND AND SEPARATIONS FOR A BIOPROCESS IS EFFECTED IN A SOLID PHASE SYSTEM TO PRODUCE THE BIOACTIVE ANTIBODY PHARMACEUTICAL AND ABLE TO BE SCALED UP AND PRODUCED INDUSTRIALLY. COLUMN PERFUSION AND ANCILLARY AUDITABLE PROCESS DATA CAN BE ARCHIVED. ACTIVATED BEADS AVAILABLE FROM BIOTECH SUPPLIERS AND SYSTEMS SUCH AS PHARMACIA BIOPILOT(GELIFESCIENCES.COM) ARE COMPUTER MANAGED AND WITH COMPATIBLE ANALYTICAL SUPPORT SYSTEMS.

Bioselective antibody conjugation is effected via a custom made biological substrate (epitope column) corresponding to the antibody recognition properties.This solid phase with biologically attached antibody forms a perfused column and a bioprocess based synthesis, separation and purification system.The bioselective antibody preparation product is derived from a managed conjugation chemistry utilizing biological specificity.

 
 

BIOSELECTIVE SOLID PHASE IMMUNOCONJUGATE SYNTHESIS

Custom biological substrate components:

Epitopes-biotechnology,peptide and glycoprotein synthesis.

Attachment to solid phase media usually in bead  form for epitope column- sequence of fluidic reagent perfusion to prepare solid phase.

Incubate with antibody fragment,separate wash and purify.

Expose to retroactivated linker reactant (eg.DTPA anhydride). Wash and purify.Chromatographic monitoring in tandem.

Solid phase media with biologically activated matrix allows automation and computer controlled conjugation bioprocess.


 

NATURE OF LABELING DAMAGE-IMPURITY

Labeling of the sensitive binding site where intimate contact is required.

Reactant chemical damage,excess and byproducts

Folding distortion particularly tertiary structure

Low retention and persistence upon binding to antigenic target

Low specific activity

Low solubility-particularly when higher substitution is attempted.

 

Clinical limitations

1.Low targeting

2.Reduced binding

3.Low potency

4.High background

5.Corrupted biodistribution

6.Side effects

7.Adverse toxicity 

 

                   ................................................................................................................


CONJUGATION BIOSELECTIVITY PROCESS

Bioselective antibody conjugation is effected via a custom made biological substrate (epitope column) corresponding to the antibody recognition properties.This solid phase with biologically attached antibody forms a perfused column and a bioprocess based synthesis,separation and purification system.The bioselective antibody preparation product is derived from a managed conjugation chemistry utilizing biological specificity.

 
 

 BIOSELECTIVE SOLID PHASE IMMUNOCONJUGATE SYNTHESIS

Custom biological substrate components:

Epitopes-biotechnology,peptide and glycoprotein synthesis.Attachment to solid phase media usually in bead  form for epitope column- sequence of fluidic reagent perfusion to prepare solid phase. Incubate with antibody fragment,separate wash and purify.Expose to retroactivated linker reactant
(eg.DTPA anhydride).Wash and purify.Chromatographic monitoring in tandem.

Solid phase media with biologically activated matrix allows automation and computer controlled conjugation bioprocess.

 

BIOSELECTIVE CONJUGATION MECHANISM

Bound antibody presents a selected conformed area for reactant access.This area is other than the binding site of the antibody.The antibody bound to the antigen has a different conformation to that of the free antibody as folding changes occur with binding.The intimate binding site molecular contact is such that every water molecule is squeezed out. Conjugation is prevented on the binding site being protected by the bound epitope. After reactant contact and washing the antibody conjugate is cleaved and separated from the solid phase by unfolding back into the aqueous phase with subsequent dose and formulation for dispensing. Conjugation reactant chemistry can be with direct or indirect linking agents.

 

ISOLATION OF ANTIBODY

 

ISOLATION OF ANTIBODY IN PREPARATION OF BIOACTIVE CONJUGATION BIOPROCESS. BOUND ANTIBODY-ANTIGEN COMPLEX ALLOWS EXPOSURE TO REACTION CHEMISTRY AND SEPARATIONS.

 
 

ANTIBODY CONJUGATION

 

REACTANT CHEMISTRY APPLIED TO BOUND ANTIBODY-ANTIGEN COMPLEX IN A SOLID PHASE ALLOWS CHOICE OF CONJUGATE TO BE ATTACHED EITHER DIRECTLY OR INDIRECTLY VIA A LINKER. CONVENTIONAL ANTICANCER DRUGS SUCH AS DOXORUBICIN AND METHOTREXATE AVAILABLE IN A PHARMACEUTICAL QUALITY ASSURED FORMULATION SOURCE AND WITH A KNOWN PHARMACOLOGICAL PROFILE. AGENTS ARE EXPECTED TO BENEFIT FROM BIOACTIVE CONJUGATION METHODOLOGY EG BR96 IMMU-11 AND MYLOTARG.

 

 

RELEASE AND SEPARATION OF PHARMACEUTICAL FROM SOLID PHASE UTILIZING REVERSIBLE BIOTECH UNFOLDING REFOLDING. FOLDING PROPERTIES PRESERVED WITH FULL BINDING SITE RECOGNITION TO PRODUCE THE BIOACTIVE IMMUNOCONJUGATE.
 

 

 

 

GLUTARALDEHYDE IS A SIMPLE VERSATILE BIFUNCTIONAL LINKING AGENT ENABLING A WIDE VARIETY OF CONJUGATES TO BE CONSTRUCTED. GLUTARALDEHYDE IS FORMULATED IN SOLUTION AND PERFUSED THROUGH COLUMN. CLEAN IN PLACE PURIFICATION AND SEPARATIONS CAN BE APPLIED.

BIOACTIVECONJUGATION TECHNOLOGY TO EHANCE TARGETING OF CYTOTOXICS UTILIZING GLUTALDEHYDE TO ACTIVATE IMMUNOGLOBULIN REACTIVITY TOWARDS -NH2 GROUPS OF ANTICANCER AGENT TO BE CONJUGATED. WITH THE ANTIBODY ATTACHED IN A STERICALLY BOUND FORM. IT IS BELIEVED EVERY WATER MOLECULE IS SQUEEZED OUT IN IMMUNOREACTION. GLUTALDEHYDE CAN ATTACH TO AREAS OTHER THAN THE BINDING SITE.TOREA BIOACTIVE ANTIBODY SYNTHESIS. INTRODUCTION OF THE CYTOTOXIC IN A SUBSEQUENT PERFUSION STEP WITH CLEAN IN PLACE WASHES WITH PHOSPATE BUFFERED SALINE AND 1% ALBUMIN.

BIOACTIVE CONJUGATION TO PRODUCE VIA SOLID PHASE BIOPROCESS THE COUPLED CYTOTOXIC WITH A FOLDED PRODUCT PURIFIED BY SEPARATIONS AND CLEAN IN PLACE STRATEGIES, RELEASE OF CONJUGATE BY UNFOLDING SEPARATES BIOPHARMACEUTICAL INTO THE AQUEOUS PHASE. SEPARATED PHARMACEUTICALREFOLDED TO REGAIN ORIGINAL TERTIARY STRUCTURE.A PROCESS EMPLOYED EXTENSIVELY IN BIOTECHNOLOGY IN RECOVERY OF BIOREACTOR PRODUCT. UREA AND GUANIDINE INTERACT WITH WATER AND HYDROPHOBIC INTERACTIONS OF THE FOLDING. A REVERSIBLE PROCESS OBSERVED WITH ENTROPY AND PATHWAY ASSEMBLY COMPATIBLE WITH TERTIARY STRUCTURE.

 

 

 

BIOACTIVE DTPA IMMUNOCONJUGATES HAVE CHELATING PROPERTIES FOR RADIOLABELING IMAGING CLINICS. GADOLINIUM IS A PARAMGNETIC AGENT USED WITH MRI IMAGING AND IS SEQUESTED BY CHELATING AGENTS SUCH AS DTPA, EVOLVED MACROCYCLES. GADOPENTATE MARKETED AS MAGNEVIST AS A VASCULARITY AGENT EXTENSIVELY IN MAGNETIC RESONANCE IMAGING CLINICS BY OMNISCAN,PROHANCE,OPTIMARK AND MULTIHANCE. YTTRIUM 90 CHELATION IS IN FOR RADIOTHERAPY WITH THE PRODUCT ZEVALIN AND IS ABLE TO BE ENHANCED IN IT'S PROPERTIES WITH BIOACTIVE CONJUGATION METHODOLOGY.

 

 

BIOACTIVE DTPA IMMUNOCONJUGATES SUITABLE FOR CHELATED LABELING WITH TECHNETIUM,YTTRIUM AND GADOLINIUM. LABELING ACHIEVES HIGH SPECIFIC MODIFICATION WHILST PRESERVING BINDING SITE RECOGNITION PROPERTIES AND FOLDING CAPABILITY. A SCALABLE AUTOMATED PROCESS ENABLES INDUSTRIAL UPTAKE AND A NUMBER OF PIPELINES.THE ANHYDRIDE OF DIETHYLENETRIAMINE PENTA-ACETIC ACID IS REACTED WITH MAB BOUND TO IT'S BIORECOGNIZED EPITOPE. THIS PRESENTS A CHELATING MIETY TO INSTANTLY LABEL EASILY AND EFFICIENTLY IN A STERILE FORMULATED VIAL KIT.


BIOSELECTIVE FOLDING IMMUNOREACTIVITY

 

It is well known that antigen binding fragments of antibodies can be refolded from denatured states with recovery of their specific binding. Renaturation and refolding is utilized extensively in separation and purification by biotechnology in product recovery from cell cultures.Guanidine and urea have been shown to reversibly unfold-refold in a highly co-operative process and their folding profiles with urea gradient concentration can be obtained. Unfolding release of immunoconjugate can be with low concs of denaturant. An immunoconjugate with folding properties results. This contributes to supramolecular and dynamic binding properties to enhance targeting ...in part the folding changes observed with binding. Binding is strong and persistent.
 
 

REFERENCES

1.Journal of Nuclear Medicine 6 27 P1040 1986

2.US Patent No 5,082,928

3.Advances in Radiopharmacology ISBN 0 642 59902 5 Int Symposium on Radiopharmacology (6th:1989 Sydney,Australia)

4.Journal of Nuclear Medicine 32 116-22 1991

5.Journal of Immunological Methods, 133 1990 159-167 
 


LUNG CANCER

Using Nebulized Biotechnology Derived Tc99m Labeled Antibodies For Early Detection Imaging

 

http://users.netaccess.co.nz/lung-cancer

CLINICAL UTILIZATION

Mucociliary Clearance   4hrs

 

    

 

 

 

Lung cancer pathology
Lung cancer is the leading form of cancer death in the US. In 1992 there were about 168,000 new cases of the disease diagnosed and approximately 146,000 deaths attributed to it. This number is increasing and many cases are not diagnosed until fairly advanced due to the rapid rate of tumor growth,soft tissue differentiation identification difficulties with cytology sampling, radiology and deep fine structure location in the bronchial tree. Location of cancers occur frequently at bronchial tree bifurcation points with aerodynamic interception of inhaled carcinogenic particulates such as tar from cigarette smoke.
Such is the burden on health care resources that tobacco litigation settlements resulted in $230 billion recently.
Current lung cancer detection
For the patient the problem has been to detect and diagnose malignant from harmless lesions and in the US 40,000 resultant invasive exploratory thoracic surgical operations(thoracotomies) requiring risky full theatre resection workup and recovery with these investigations are found to be inappropriate for them. In these cases the bronchial wall lesions can be benign resulting from other etiology e.g. inflammation,infection and fibrosis or part of unresectable disease and disseminated. Other options for inoperable disease that has spread include chemotherapy and palliative measures.
New imaging technology
A new imaging technology has been developed in Dunedin New Zealand to photograph epithelial lung cancer by inhalation delivery and localization of a biospecific contrast  tracer. The procedure is simple and non-invasive involving breathing of a mist and offers imaging with cancer for early and more specific detailed photography available at hospital imaging centres on an outpatient basis using currently available ethically approved clinical materials and procedures. The inhaled biospecific contrast agent is able to recognize epithelial lung cancer and localize by antibody reaction carrying the imaging agent with it once inside the lungs and deposited on the lung epithelium. The resultant images can be used in treatment planning eg radiotherapy targeting from early diagnosis and screening.

Inhalation of the biospecific mist is an efficient direct delivery means achieved with nebulizer produced aerosol having microfine droplet dimensions to achieve deep lung penetration. Micromist droplet size must be less than 3 microns. Droplets with greater diameter deposit in the upper airways(eg throat) with participation in air filtration for breathing. Coating of the considerable surface of the bronchial tree occurs beyond the range of the bronchoscope in depth and spatially within airways architecture.
 

Biospecificity of monoclonal antibodies
Differentiating malignant from innocent lesions is achieved by the monoclonal antibody based tracer which has fine tuned recognition properties based on a molecular level. Antibodies are utilized in the body as a protection and recognition means e.g. flus and immunity and have offered optimism in oncology since their discovery and Nobel Prize winning biotechnological development. Application of these is widespread and cytology of tumor tissue identification in sputum and biopsy samples is used extensively in bioseparation technologies. Monoclonals and recognition derivatives are now available as pharmaceutical products with high purity and in quantity from the biotech industry and stock market investment. Labeling and imaging utilizes the clinical tracer Technetium used extensively in hospital nuclear medicine clinics and nebulizer produced aerosol for routine ventilation studies .

Epithelial lung cancers have tumor specific characteristics and the antibodies selected are able to recognize these and  immunoreact. Contrast enhancement then occurs with mucociliary clearance sweeping out the unreacted agent from the lung fields and background subtraction for the image process. This natural clearance develops biologically based imaging photographs showing location and extent of the cancer. Mucociliary clearance in participation with lung fine structure acts as a powerful filtration mechanism for our inhaled air.

 

 

 		 

CHEMOTHERAPY

 

Cytotoxic Therapy Augmentation with bioselective immunoconjugates

 

PRODUCTION OF A TARGETED CYTOTOXIC. BIOACTIVE IMMUNOCONJUGATE CYTOTOXIC PHARMACEUTICAL. LINKAGE CHEMISTRY.APPLICATION OF REACTANT CHEMISTRY TO ANTIBODY IN EXPOSED TOPOLOGY OF THE MOLECULAR REGION OTHER THAN THE BINDING SITE AND IN THE BOUND FOLDED FORM.WITH UNFOLDING SEPARATION OCCURS FROM THE SOLID PHASE. REVERSIBLE REFOLDING ALLOWS FULL BINDING CAPACITY AND PERSISTENCE.

Cytotoxics with more specifically targeted properties- therapeutic index increased with biospecificity.

Useful particularly in cases of motile and diffuse cancers such as leukaemia and occult metastatic deposition.

Increase of dosage-Tolerance of a higher dosage of cytotoxic to be administered with greater concentration at tumor sites increasing oncological kill.

Reduction of side effects-Cytotoxic side effects of current chemotherapy severe-Often from non specificity,cross reactivity and compromised biodistribution. Frequently limits treatment plan eg nausea,wbc and haematological,hair loss,cardiotoxicity and myelosuppression side effects.

CYTOTOXICS OF INTEREST

 

Bioselective immunoconjugate cytotoxic composites eg methotrexate,doxorubicin

Established mode of action-Antimetabolite,alkylating etc

Widely used,..extensive clinical preparations of source available to pharmaceutical quality.

Known biodistribution,dosage and antineoplastic mode of action.

Toxicological data from pharmacological and pharmacokinetic studies.
 

BIOPROCESS PATHWAY FOR PRODUCTION OF MONOCLONAL BASED BIOACTIVE PHARMACEUTICALS. CYTOTOXIC PREPARATION WITH SEQUENCE OF REACTION AND PURIFICATION STEPS WITH PERFUSION AND CONTROL IN A COMPUTER COLUMN APPLICATION SUCH AS BIOPILOT. SOLID PHASE MEDIA OF THE IMMUNOADSORBENT BEADS ALLOWS HIGH SPECIFIC ATTACHMENT WITH PROTECTED BINDING SITE AND FOLDING CAPABILILITY FREE FROM NON IMMUNOCOMPETENT IMPURITIES. HIGH SPECIFICITY FOR A FAVOURABLE TARGET TO BACKGROUND RATIO WHICH INCREASES EFFICACY.
 
 
 

BIOSELECTIVE CYTOTOXIC LINKAGE CHEMISTRY

DIRECT....methotrexate anhydride,activated esters,retroactivated moieties.

INDIRECT....Linkers..sdtp,glutaraldehyde

Delivery Toxins.... ricin,gelonins,tricosanthins,tricokerin,pokeweed,diptheria and pseudomonas toxins.Cytokines eg interleukin 2

BIOSELECTIVE CYTOTOXIC PRODUCTION

 

Preparitive Conjugation Bioprocess

Biotech Compatible

Hardware..columns,valves,conduit and flow circuits eg Pharmacia,Biorad

Tandem chromatographic support eg detection and analysis equipment...uv spectroscopy

Reaction and sequence if automated...liquids,reagents,origins and batch

Software archive..script design eg biopilot...commercially supplied

Site...cleanroom suite

Records available for independent regulatory inspections. Batch requirements...Standard compliance validation...audit trail.
 

BIOSELECTIVE IMMUNOCONJUGATE SYNTHETIC STEPS

 

Column perfusion sequencing

1.Attach antigen.

 

DIAGRAM OF BIOACTIVE ANTIBODY PREPARATION BIOPROCESS OF SOLID PHASE MEDIA IMMUNOADSORBENT WITH ANTIGEN COUPLING AND CHEMISTRY. ATTACHMENT TO AGAROSE/ACRYLAMIDE BEADS ACTIVATED WITH ESTERS OR ANHYDRIDES OBTAINED COMMERCIALLY(EUPERGIT C OXIRANE,N-HYDROXYSUCCINIMIDE NHS SEPHAROSE. COUPLING OF ANTIGEN TO MEDIA IS ROUTINE RELIABLE BIOCHEMICAL PROCEDURE USED IN PROTEIN PURIFICATION.

REACTION MONITORING OF ANTIGEN EPITOPE ATTACHMENT TO SOLID PHASE MEDIA FOR BIOPILOT BIOPROCESS IN PRODUCTION OF BIOACTIVE ANTIBODY CONJUGATES. UV SPECTROSCOPY IS ABLE TO QUANTITATIVELY DETERMINE CONCENTRATION LINKED TO ACTIVATED SEPHAROSE COLUMN AND UNDER AUTOMATED COMPUTER CONTROL AND ARCHIVE. EPITOPES ARE AVAILABLE FROM SOURCES IN BIOTECHNOLOGY AND SPECIALIZED CHEMISTRY SYSTEMS SUCH AS PEPTIDE SYNTHESIS.

2.Allow antibody attachment.

ISOLATION OF ANTIBODY FOR EXPOSURE TO CONJUGATION CHEMISTRY IN BIOPROCESS OF PRODUCTION OF A BIOACTIVE IMMUNOCONJUGATE. BOUND ANTIGEN PROTECTS THE BINDING SITE OF IMMUNOGLOBULIN WITH THE CLOSE CONTACT OF THE IMMUNO STERIC FIT FROM REACTANTS WITH THIS REACTION OCCURING WITH THE ANTIBODY IN IT'S FINAL FOLDED CONFORMATION. THIS IS DIFFERENT TO CONJUGATION APPLIED TO A FREE FORM ANTIBODY IN SOLUTION.(TERTIARY STRUCTURE)

REACTION MONITORING OF ANTIBODY OR FRAGMENT ATTACHMENT TO SOLID PHASE MEDIA FOR BIOPILOT BIOPROCESS IN PRODUCTION OF BIOACTIVE IMMUNOCONJUGATES. ANALYSIS OF INLET AND EXHAUST CONCENTRATION IS ACHIEVED WITH ULRA VIOLET ABSORBANCE AT 280NM. GELIFESCIENCES SUPPLIES HIGH GRADE MEDIA AND CONTROL SYSTEMS FOR THE BIOPROESS. ANTIBODIES AND FRAGMENTS ARE AVAILABLE FROM BIOTECHNOLOGY WITH RECOMBINANT CULTURE AND FERMENTATION TECHNOLOGY.

3.Expose to conjugation chemistry.

4.Release of antibody conjugate by unfolding.

5.Refold by salt extraction...dry gel filtration,dialysis,cross flow tangential...Amicon.

6.Attach subsequent moiety as required eg contrast agent
 

Automated with fluidic feed porting for downstream process with tandem chromatographic support.

Each purification stage allows washing and separation

FLUIDIC CONTROL IN BIOACTIVE ANTIBODY SYNTHETIC BIOPROCESS PRODUCTION. MEDIA BED OF ACTIVATED GEL BEADS AS SUPPORT TO ALLOW SOLID PHASE EFFICIENCIES WITH EFFECTIVE SEPARATIONS OF REACTANT EXCESS.BYPRODUCTS AND IMMUNOIMPURITIES. SOLID PHASE TO DRIVE REACTIONS TO COMPLETION WITH TOLERANCE TO HIGHER REACTION CONCENTRATIONS WITH GREATER YIELDS. AVAILABLE TO GMP AND QUALITY CONTROL PERTAINING TO PHARMACEUTICAL USE.BINDING SITE PROTECTED AND FOLDING CAPABILITY FOR SPECIFICITY AND PERSISTENCE OF IMMUNOREACTION.
 
 
 
 SCANNING ELECTRON MICROSCOPE IMAGE OF BIOPROCESS AFFINITY MEDIA INCORPORATED INTO PRODUCTION OF BIOACTIVE ANTIBODY CONJUGATES. SEPHAROSE BEADS ACTIVATED TO FORM A SOLD PHASE IMMUNOADSORBENT BED INTERACT WITH A LIQUID REACTANT PHASE FOR PURIFICATIONS AND SEPARATIONS. IMMUNOREACTIVITY ASSAY

IMMUNOREACTIVITY ASSAY PROCEDURE FOR BIOACTIVE ANTIBODY.COMPETITIVE UPTAKE IN A SOLID PHASE DETECTION SYSTEM. ANTIBODY FOLLOWS IN VIVO LOCALISATION PATTERN WITH 98 PERCENT IMMUNOREACTIVITY UPTAKE WITHIN 48HRS.

 


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IMAGING

 

Advantages of bioselective scanning immunoconjugates

 

Diagnostic imaging contrast agents

 

SCINTIGRAPHY
MRI

Clinical high resolution imaging from enhanced bioactive targeting properties of agents with increased tumor uptake and lower background. Higher specific activity of agents whilst retaining immunoreactivity and binding. Earlier more accurate diagnosis and staging- quicker image acquisition time.

SCINTIGRAPHY

Clinically utilized in non-invasive screening -Hospital Nuclear Medicine Departments-gamma camera imaging

Alternative generic linked and chelated radionuclides Tc 99m I123 In111

Diverse antigenic targets eg fibrin,myosin and cancer

Higher substitution of bifunctional chelators and linkers eg.dtpa,dota,mag,bat, macrocycles and biotin.

Label attachment for activated antibody eg Tc99m from generator eg stannous reduction

Availability as kit ready to use from conjugation bioprocess

BIOACTIVE TECHNETIUM 99M ISOTOPE LABELING OF A MONOCLONAL ANTIBODY ABLE TO BE ROUTINELY USED IN NUCLEAR MEDICINE CLINICS.TECHNETIUM IS LABELED VIA DTPA AND IS SUITABLE FOR HUMAN USE WITH PHARMACEUTICAL QUALITY,SPECIFICATIONS,DOSAGE AND USE WITH DIAGNOSTIC IMAGING.

TEHNETIUM BIOACTIVE IMMUNOCONJUGATES PREPARED IN A BIOPROCESS. FULL BINDING SITE RECOGNITION AND NATIVE FOLDING CAPACITY ALLOW HIGHER SPECIFIC LABELING AND USE FOR EARLY NON INVASIVE DIAGNOSIS WITH BETTER IMAGE RESOLUTION. IMMUNOREACTIVE AND PERISTENT ANTIBODY LOCALISATION ATTRIBUTES ALLOW HIGHER TARGET TO BACKGROUND RATIOS.

 

 

MRI 

Magnetic resonance imaging

Gadolinium MRI scanning

GADOLINIUM IS A PARAMAGNETIC MRI CONTRAST AGENT ABLE TO BE ATTACHED TO ACHIEVE GREATER AND EARLIER DETAIL IN MAGNETIC RESONANCE IMAGING. BIOACTIVE CONJUGATION ALLOWS CHELATION WITH HIGH SPECIFICITY ALLOWING HIGH RESOLUTION AT AREAS OF CANCER, FASTER IMAGING AQUISITION TIME

Magnetic contrast agents linked via bifunctional linkers eg gadopentate

Greater potency ensuring better tumor uptake increasing magnetic field strength at lesion site.

Alternative generic chelated paramagnetic agents becoming available eg Mn

Availability as kit ready to use from bioprocess

RADIOTHERAPY

Therapeutic Radionuclide Labels

Yttrium, Copper, Samarium via Chelated Linkers

eg. DTPA, DOTA, MAG, MACROCYCLES

Yttrium-90 Ibritumomab Zevalin is available commercially

Efficacy is able to be enhanced with bioactive attachment

Conjugated

eg: Iodine 125, 131 Via Bolton Hunter

Iodine-131 tositumomab Bexxar is available commercially

Efficacy is able to be enhanced with bioactive attachment

 

                      

ISOTOPES OF IODINE ARE AVAILABLE AND OF VALUE IN IMAGING I123 AND RADIOTHERAPY II31. APPLICATIONS FROM BIOACTIVE CONJUGATION ALLOWS HIGH SPECIFIC LABELING TO BE ACHIEVED MAINTAINING BINDING SITE RECOGNITION, NATIVE FOLDING AND INCREASE IN EFFICACY.IODINE-131 BEXXAR TOSITUMOMAB IS AVAILABLE CLINICALLY AND IS EXPECTED TO BENEFIT FROM BIOACTIVE LABELING

 

Attractive due to potency and small size for micrometasases, dissemination,conventional drug resistance and motile cancers

Alpha, Beta emitters with Dosimetry
 

                             .................................................................................
  REFERENCES FOR TOREA BIOACTIVE ANTIBODY CONJUGATION
  RESEARCH JOURNAL OF NUCLEAR MEDICINE ADELSTEIN ET AL HARVARD MEDICAL SCHOOL BINDING SITE PROTECTION LEADS TO HIGHER IMMUNOREACTIVE FRACTION.

RESEARCH METHOD OF PREPARING CONJUGATED ANTIBODIES US PATENT 5082928

 

 

 

Torea Technologies has applications in these areas and seeks to develop and utilize the Technologies in view of their compassionate benefits in cancer

 

Torea Technologies

 Mark P Best

     PO Box 823

     Dunedin

     Aotearoa New Zealand

     e-mail:torea@netaccess.co.nz

 

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