Wednesday, December 28, 2016

SHIKARI® S-ATA v2 ELISA kit for quantification of anti drug antibodies to ADALIMUMAB (HUMIRA®) with confirmation



Matriks Biotek® to introduce SHIKARI® S-ATA v2 ELISA kit to measure anti-adalimumab antibody levels and confirmation of the results in the same kit. 

Adalimumab (Humira®) is a fully humanized monoclonal antibody against TNF-α, and is approved for the treatment of RA and Crohn's disease (CD).  Even though it is fully humanized, adalimumab does not eliminate the risk of immunogenicity in both CD and RA patients.

Generation of antibodies-to-adalimumab (ATA) in the serum is associated with lower serum adalimumab concentrations and reduced response rate to treatment. 

 SHIKARI® Q-ADA v1 kit together with SHIKARI® S-ATA v2 ELISA kit can give definitive information about patient status when evaluated with different tests and clinical status of the patient. By knowing the trough level of the drug e.g. if it is below the Cmin of the drug and there is anti-drug antibodies then the course of treatment should be changed accordingly. Adalimumab dose can be escalated, other drugs can be added to treatment and end up with switching to another biological if necessary. By following patient’s adalimumab and anti-adalimumab drug (ATA) levels. Methotrexate or leflunomide prevents the development of neutralizing Abs against adalimumab so knowing and following up of anti-adalimumab drug levels will be highly valuable in the course of treatment. 

One of the major concern when detecting anti-adalimumab drug (ATA) levels is false positivity of the results. SHIKARI® S-ATA v2 ELISA kit can detect 30 ng/ml of ATA and to eliminate false positivity optimized Confirmation Regeant is added to the test system for convenience. 

You can find related and all 38 peer reviewed publications done by SHIKARI® ELISA kits at http://matriksbiotek.blogspot.com.tr/2016/12/publications-done-by-using-shikari.html

For product insert please refer to 
http://matriksbiotek.com/files/manuals/S-ATA.pdf





Monday, December 19, 2016

SHIKARI® S-AIR V2 ELISA kit for Quantification of anti drug antibodies to INFLIXIMAB BIOSIMILAR CT-P13 (Remsima®; Inflectra®) with confirmation


A biosimilar is highly similar to, but not exactly the same as the existing, FDA-approved biologic, called the “reference” drug. People are familiar with generic versions of brand-name drugs, but biosimilars are not generic drugs. Generic versions of brand-name drugs are exact copies of chemically synthesized medicines. Biosimilars are not-quite-perfect copies of biologics drugs derived from living cells that are impossible to replicate exactly.

Development of biosimilar proteins will present drug developers with a variety of challenges, both in manufacturing and in the clinic. Immunogenicity will prove to be one of the biggest challenges. Proper design and validation of an assay to detect anti-drug antibodies and accurate interpretation of sample analysis results will prove integral to developing a biosimilar protein.

Taking the above and below-mentioned into account Matriks Biotek® produced SHIKARI® S-AIR v2 for quantitative ELISA kit as the  first ELISA kit for the biosimilar of Infliximab, CT-P13 (Remsima®; Inflektra®) with confirmation reagent. 

Comparisons for measuring especially ADA’s and trough levels are made by the tests developed for reference drug, not for biosimilar which may (or may not) differ on the test results. On the contrary we do not know the results when comparisons done by the ELISA kit developed for the biosimilar drug, it can present low immunogenicity as well. Regulatory organizations (FDA, EMEA) requires PK/ADA tests to be done by biosimilar drug in comparison to reference drug.

Matriks Biotek® offers to develop custom ELISA kits for your biosimilars or biologics on contract manufacturing basis depending on the molecule provided and with the agreements done by other CRO and GLP companies can also provide Phase 1 studies in collaboration. 

Matriks Biotek® is the first company to commercialize SHIKARI® ELISA kits for biological drug testing to measure trough levels and anti-drug antibodies (immunogenicity) since 2008.

For product insert:
http://matriksbiotek.com/files/manuals/S-AIR.pdf

REFERENCES:
1.       Farkas K., et al., Efficacy of infliximab biosimilar CT-P13 induction therapy on mucosal healing in ulcerative colitis. Journal of Crohn's and Colitis Advance Access published April 21, 2016
2.       Malíckova K, et al., Serum trough infliximab levels: A comparison of three different immunoassays for the monitoring of CT-P13 (infliximab) treatment in patients with inflammatory bowel disease, Biologicals 2015.
3.       Farkas K. et al, Efficacy of the new infliximab biosimilar CT-P13 induction therapy in Crohn’s disease and ulcerative colitis experiences from a single center. Expert Opin. Biol. Ther. 15:(9) 2015.


Thursday, December 15, 2016

Matriks Biotek® to introduce SHIKARI® Q-ATI v6 ELISA kit to measure anti-infliximab antibody levels and confirmation of the results in the same kit.



Infliximab (TNF alpha blocker) has improved the treatment of inflammatory diseases such as inflammatory bowel disease and rheumatoid arthritis. Some of the patients do not respond to induction therapy, and during the time course of treatment secondary loss of response may occur and these percentages may reach to 50% of patients under treatment.

By following patient’s infliximab and ant-infliximab drug (ADA) levels can give definitive information about patient status when evaluated with different tests and clinical status of the patient. By knowing the trough level of the drug e.g. if it is below the Cmin of the drug and there is anti-drug antibodies then the course of treatment should be changed accordingly. Infliximab dose can be escalated, other drugs can be added to treatment and end up with switching to another biological if necessary.


Precise measurement trough and ADA levels of infliximab gained high importance. Knowing anti-infliximab levels of the patient along with the confirmation of positivity to prove the reaction is not false positive. Peer reviewed publications cumulated since Matriks Biotek® first commercialized SHIKARI® ELISA kits in the whole world market at 2008 
(http://matriksbiotek.blogspot.com.tr/2016/12/publications-done-by-using-shikari.html)
For product insert:
http://matriksbiotek.com/files/manuals/Q-ATI.pdf

Tuesday, December 13, 2016

Two research articles added to the list of publications done by using Q-BEVA SHIKARI® ELISA kits : For precise measurement bevacizumab from different samples.


Biological activities of the rice-derived Bevacizumab from transgenic rice callus and from the plasma of mice were detected using by Q-BEVA SHIKARI® ELISA kit.





1.    Chen L., et al., Efficient Production of a Bioactive Bevacizumab Monoclonal Antibody Using the 2A Self-cleavage Peptide in Transgenic Rice Callus. Frontiers in Plant Science August 2016 | Volume 7 | Article 1156.
     http://journal.frontiersin.org/article/10.3389/fpls.2016.01156/full

2.  Bergen T.V., et al., Complementary effects of bevacizumab and MMC in the improvement of surgical outcome after glaucoma filtration surgery. Acta Ophthalmologica 2015, 667-678.
     https://www.ncbi.nlm.nih.gov/pubmed/25988844 


For further information and pricing please contact us from info@matriksbiotek.com


Thursday, December 8, 2016

Publications done by using SHIKARI® ELISA kits from Matriks Biotek® updated 06.02.2017: List of 40 peer reviewed journal articles and 10 poster presentations



PATENT:
1.    CONTROLED DRUG DELIVERY SYSTEMS FOR ANTI-TNF-α
WO 2014046631 A1

PhD Thesis:
1.    EVALUATION of TRANSSCLERAL PERMEABILITY EFFECTS of NANOTECHNOLOGY BASED DRUG DELIVERY SYSTEM in OCULAR DRUG APPLICATIONS


Dr. NAGİHAN UĞURLU
Hacettepe University
In Turkish with English abstract.

PEER REVIEWED JOURNAL ARTICLES:

1.    Gibellini L, De Biasi S, Bianchini E, et al. Anti-TNF-α Drugs Differently Affect the TNFα-sTNFR System and Monocyte Subsets in Patients with Psoriasis. Richard Y, ed. PLoS ONE. 11(12), 2016.
2.    Julsgaard M, Christensen LA, Gibson PR, Gearry RB, Fallingborg J, Hvas CL,Bibby BM, Uldbjerg N, Connell WR, Rosella O, Grosen A, Brown SJ, Kjeldsen J,Wildt S, Svenningsen L, Sparrow MP, Walsh A, Connor SJ, Radford-Smith G, LawranceIC, Andrews JM, Ellard K, Bell SJ. Concentrations of Adalimumab and Infliximab in Mothers and Newborns, and Effects on Infection. Gastroenterology. Jul;151(1):110-9, 2016.
3.    Jonathan GG, Beatriz AÁ, Nazco C GJ, Norberto BL, Fernando GN. Plasma levels of trastuzumab in gastric cancer: Case report. J Oncol Pharm Pract. Sep 23, 2016.
4.    Choi SY, Kang B, Lee JH, Choe YH. Clinical Use of Measuring Trough Levels and Antibodies against Infliximab in Patients with Pediatric Inflammatory Bowel Disease. Gut Liver. Sep 9 2016.
5.    Chen L., et al., Efficient Production of a Bioactive Bevacizumab Monoclonal Antibody Using the 2A Self-cleavage Peptide in Transgenic Rice Callus. Frontiers in Plant Science August 2016 | Volume 7 | Article 1156.
6.    Farkas K., et al., Efficacy of infliximab biosimilar CT-P13 induction therapy on mucosal healing in ulcerative colitis. Journal of Crohn's and Colitis Advance Access published April 21, 2016
7.    Gutiérrez A, Zapater P, Juanola O, Sempere L, García M, Laveda R, Martínez A, Scharl M, González-Navajas JM, Such J, Wiest R, Rogler G, Francés R. Gut Bacterial DNA Translocation is an Independent Risk Factor of Flare at Short Term in Patients with Crohn's Disease. Am J Gastroenterol. Apr;111(4):529-40, 2016
8.    Won Jae Song, Ben Kang, So Yoon Choi, and Yon Ho Choe. Adalimumab Treatment in Pediatric-Onset Crohn’s Disease Patients after Infliximab Failure: A Single Center Study. Pediatr Gastroenterol Hepatol Nutr. Jun; 19(2): 116–122, 2016.
9.    Hayashi S, et al., Early Prognostic Factors Associated with the Efficacy of Infliximab Treatment for Patients with Rheumatoid Arthritis with Inadequate Response to Methotrexate. Rheumatol Ther (2016) 3:155–166
10.  Bortlik M., et al., Discontinuation of anti-tumor necrosis factor therapy in inflammatory bowel disease patients: a prospective observation. Scandinavian Journal of Gastroenterology, 2016 vol. 51, no. 2, 196–202.
11.  Al-Karkhi M.A., et al., Correlation between Anti-infliximab and Anti-CCP Antibodies Development in Patients with Rheumatoid Arthritis Treated with Infliximab in Baghdad Teaching Hospital. IOSR Journal of Dental and Medical Sciences, Volume 14, Issue 11 Ver. IV (Nov. 2015), PP 95-100.
12.  Kui R, Gál B, Gaál M, Kiss M, Kemény L1, Gyulai R. Presence of antidrug antibodies correlates inversely with the plasma tumor necrosis factor (TNF)-α level and the efficacy of TNF-inhibitor therapy in psoriasis. J Dermatol. Sep;43(9):1018-23, 2016
13.  Bodini G., Edoardo G. Giannini, Edoardo V. Savarino, and Vincenzo Savarino. Adalimumab Trough Levels and Response to Biological Treatment in Patients With Infl ammatory Bowel Disease: A Useful Cutoff in Clinical Practice. Am J Gastroenterol. Vol. 110, March, 2015.
14.  Juanola O., et al., Anti-TNF-alpha loss of response is associated with a decreased percentage of FoxP3+ T cells and a variant NOD2 genotype in patients with Crohn’s disease. J Gastroenterol (2015) 50:758–768.
15.  Fujita T, Murata Y, Hemmi S, Kajiwara M, Yabuki M, et al. (2015) Persistent Complement Activation is Associated with Insulin Resistance and Chronic Inflammation in Overweight Patients with Type 2 Diabetes with Dyslipidemia. Int J Immunol Immunother 2:007, 2015
16.  Malíckova K, et al., Serum trough infliximab levels: A comparison of three different immunoassays for the monitoring of CT-P13 (infliximab) treatment in patients with inflammatory bowel disease, Biologicals 2015.
17.  Farkas K. et al, Efficacy of the new infliximab biosimilar CT-P13 induction therapy in Crohn’s disease and ulcerative colitis experiences from a single center. Expert Opin. Biol. Ther. 15:(9) 2015.  
18.  Lee Y.M. et al, Infliximab ‘‘Top-Down’’ Strategy is Superior to ‘‘Step-Up’’ in Maintaining Long-Term Remission in the Treatment of Pediatric Crohn Disease. JPGN 60: (737–743), 2015
19.  Al-Karkhi M.A., et al., Development of Antibodies against Infliximab in Iraqi Patients with Rheumatoid Arthritis. J Fac Med Baghdad, 57: (241-243), 2015.
20.  Bergen T.V., et al., Complementary effects of bevacizumab and MMC in the improvement of surgical outcome after glaucoma filtration surgery. Acta Ophthalmologica 2015, 667-678.
21.  Aydın C, Ataoğlu H., Demonstration of β-1,2 Mannan Structures Expressed on the Cell Wall of Candida albicans Yeast Form But Not on the Hyphal Form by Using Monoclonal Antibodies Mikrobiyol Bul 49(1): 66-76, 2015.
22.  Pallagi-Kunstár É. et al., Utility of serum TNF-a, infliximab trough level, and antibody titers in inflammatory bowel disease. World J Gastroenterol. 20(17): (5031-5035), 2014.
23.  Khanna R., et al., Therapeutic Drug Monitoring of TNF Antagonists in Inflammatory Bowel Disease. Gastroenterology & Hepatology, August (478-489),2014.
24.  Krajcovicova A. et al., Delayed hypersensitivity reaction after initial dose of infliximab: a case report. European Journal of Gastroenterology& Hepatology 26:(485-487), 2014.
25.  Erdemli Ö., et al, In vitro evaluation of effects of sustained anti-TNF release from MPEG-PCL-MPEG and PCL microspheres on human rheumatoid arthritis synoviocytes.2014. Journal of Biomaterials Application, 29(4):524-42, 2014         
26.  Avdeeva A.A., Aleksandrova E.N., Karateev D.E., Luchikhina E.L., Novikov A.A., Cherkasova M.V., Nasonov E.L. EFFICACY OF ADALIMUMAB IN EARLY RHEUMATOID ARTHRITIS IN RELATION TO ITS SERUM LEVEL AND THE PRESENCE OF ANTI-DRUG ANTIBODY. Rheumatology Science and Practice. 52(6):624–630.  2014 Russian, English abstract http://rsp.ima-press.net/rsp/article/view/2007
27.  Bortlik M, et al, Impact of Anti-Tumor Necrosis Factor Alpha Antibodies Administered to Pregnant Women With Inflammatory Bowel Disease on Long-term Outcome of Exposed Children. Inflamm Bowel Dis 20 : (495-451), 2014.
28.  Jung Y et al., “Temperature-modulated noncovalent interaction controllable complex for the long-term delivery of etanercept to treat rheumatoid arthritis”, J. Control. Release 2013;
29.  Gutierrez A, et al, Genetic susceptibility to increased bacterial translocation influences the response to biological therapy in patients with Crohn’s disease, Gut 0:1–9, 2013.
30.  Grosen A., et al, Infliximab concentrations in the milk of nursing mothers with inflammatory bowel disease, J Crohns Colitis 2013.
31.  Romero G., et al, Poly(Lactide-co-Glycolide) Nanoparticles, Layer by Layer Engineered for the Sustainable Delivery of AntiTNF-α. Macromol. Biosci. 13: (903–912), 2013.       
32.  Cheong C, et al, Etanercept Attenuates Traumatic Brain Injury in Rats by Reducing Brain TNF-α Contents and by Stimulating Newly Formed Neurogenesis, Mediators of Inflammation, 2013; Volume 2013, Article ID 620837, 9 pages http://dx.doi.org/10.1155/2013/620837
33.  Bortlik M et al, “Pregnancy and newborn outcome of mothers with inflammatory bowel diseases exposed to anti-TNF-a therapy during pregnancy: three-center study”, Scandinavian Journal of Gastroenterology. 48: 951–958, 2013
34.  Bortlik M, et al, Infliximab trough levels may predict sustained response to infliximab in patients with Crohn's disease, Journal of Crohn's and Colitis 2012.
35.  Malickova K, et al, Phosphatidylserine-dependent anti-prothrombin antibodies (aPS/PT) in infliximab-treated patients with inflammatory bowel diseases, Autoimmun Highlights, 2012
36.  Takahashi H, et al, Plasma trough levels of adalimumab and infliximab in terms of clinical efficacy during the treatment of psoriasis, Journal of Dermatology 2012; 39: 1- 4.
37.  Seok Lee Y, et al, “Efficacy of Early Infliximab Treatment for Pediatric Crohn’s Disease: A Three-year Follow-up”, Pediatric Gastroenterology, Hepatology & Nutrition 2012; 15(4):243-249
38.  Molnar T, et al, “Importance of trough levels and antibody titers on the efficacy and safety of Infliximab therapy in inflammatory bowel disease”, Z Gastroenterol 2012; 50 - A53
39.  Kato S, et al, “Elevated Serum IgE Prior to Acute Severe Infusion Reaction During Infliximab Maintenance Therapy in a Crohn’s Disease Patient”, Crohn’s & Colitis Foundation of America 2011

40.  Adisen E, et al, “Anti-infliximab antibody status and its relation to clinical response in psoriatic patients”: A pilot study, Journal of Dermatology 2010; 37: 708–713.
POSTER PRESENTATIONS
1.    Malickova K, et al,. Monitoring patients treated with infliximab: Assessing Anti-Infliximab antibodies, Czech Republic. EUROMEDLAB. Autoimmune diseases 2009. S115       
2.    Malickova K, et al, Formation of antiphospholipid antibodies and antibodies to infliximab in anti-TNF-alpha antibody-treated patients with inflammatory bowel diseases, Charles University in Prague Czech Republic (2011)  
3.    Malickova K, et al, Relationship between serum trough infliximab levels, serum antibodies to infliximab, serum albumin levels and clinical response to infliximab treatment in patients with inflammatory bowel diseases, Charles University in Prague Czech Republic. (2011) 
4.    Lukas M, et al, Anti-infliximab antibodies in routine clinical practice is it worth to assess them, Czech Republic   
5.    Duricova D, et al, Predictors of sustained response to infliximab with Crohn’s disease: A single cohort study, Czech Republic   
6.    Bodini G, et al, Correlation between Adalimumab trough serum concentration, Anti-Adalimumab antibodies and TNF-Alpha levels with clinical outcome in patients affected by Crohn’s disease,. United European Gastroenterology. Italy 2013.       
7.    Julsgaard M, et al, Time since last drug exposure in pregnancy determines Adalimumab and Infliximab levels in neonates(Era Study), Italy 2014
8.    Szepes Z., et al, Clinical utility of measuring serum TNF alpha level, anti TNF alpha levels and antibody titers in critical situations in inflammatory bowel disease and in psoriasis, ECCO 2014 Inflammatory Bowel Disease.
9.    Julsgaard M., et al, Intra-uterine Exposure to Anti-TNF-alpha therapy(ERA study):Infliximab and adalimumab cord blood levels correlate with maternal levels at birth, ECCO 2014 Inflammatory Bowel Disease.

10.  Goldberg R., et al, Predictors of sub-therapeutic infliximab oradalimumab trough levels and anti-drug antibodies and their influence on therapeutic decisions, ECCO 2014 Inflammatory Bowel Disease.

Saturday, June 18, 2016

The first commercial ELISA kits optimized for detecting infliximab biosimilar Remsima® levels and immunogenicity for the first biosimilar


Matriks Biotek® produced SHIKARI® Q-REMS and S-AIR as the  first ELISA kits for the biosimilar of Infliximab, CT-P13 (Remsima®)and now also offers to develop custom ELISA kits for your biosimilars or biologics on contract manufacturing basis depending on the molecule provided. Matriks Biotek® is the first company to commercialize SHIKARI® ELISA kits for biological drug testing to measure trough levels and anti drug antibodies (immunogenicity) since 2008. 


Pricing for SHIKARI® Q-REMS and S-AIR is 450 Euro plus shipment per kit for a limited time until the end of September 2016. 

Do not forget to add code "Q-REMS" in your purchase order in your e-mail to info@matriksbiotek.com  

Ask for all 23 SHIKARI®  ELISA kits for biological drug testing or for your custom needs ..


Sunday, June 5, 2016

First commercial SHIKARI® kits for nivolumab (Opdivo®) in the whole world marketfrom Matriks Biotek®




By the mid of June with these new additions there will be total of 24 kits for biological drug testing for 12 different drugs widely used for treatment of diseases, such as inflammation, cancer and allergic disorders and osteoporosis.
As with all therapeutic proteins, there is a potential for immunogenicity. In order to benefit from your most valuable biological drug we recommend to measure trough levels along with anti-drug antibodies together from samples taken just before the administration of following dose by SHIKARI® ELISA kits.
We are hoping that patients, clinicians and biosimilar drug producers benefit from our products at most. Others to come soon...
Nivolumab is a humanized IgG4 monoclonal anti-PD-1 antibody. Its molecular weight is 146 kDa. Mechanism of Action: Programmed death 1 (PD-1) receptor is a type I membrane protein of 268 amino acids. PD-1 is expressed on the surface of activated T cells, B cells, and macrophages. The binding of PD-1 and its ligands (PD-L1 and PD-L2) on a tumor cell contributes to inhibition of active T-cell immune surveillance of tumors. By inhibiting the PD-1 receptor from binding to its ligands, nivolumab reactivates tumor-specific cytotoxic T lymphocytes in the tumor microenvironment and reactivates anti-tumor immunity. Clinical Dose Selection: The selection of nivolumab dose and schedule of 3 mg/kg Q2W was based on the observed clinical safety and efficacy from 306 patients in trial MDX1106-03 across different dose levels and tumor types.
Immunogenicity:
As with all therapeutic proteins, there is a potential for immunogenicity. Of 532 patients who were treated with OPDIVO 3 mg/kg every 2 weeks and evaluable for the presence of anti-nivolumab antibodies, 67 patients (12.6%) tested positive for treatmentemergent anti-nivolumab antibodies by an electrochemiluminescent (ECL) assay. Neutralizing antibodies against nivolumab were detected in five patients (0.9%). There was no evidence of altered pharmacokinetic profile or toxicity profile with anti-nivolumab binding antibody development. Of 105 patients who were treated with OPDIVO in combination with ipilimumab and evaluable for the presence of anti-nivolumab antibodies, 23 patients (21.9%) tested positive for treatment-emergent anti-nivolumab antibodies by an ECL assay. Neutralizing antibodies against nivolumab were detected in one patient (1%). There was no evidence of altered toxicity profile with anti-nivolumab antibody development. The detection of antibody formation is highly dependent on the sensitivity and specificity of the assay.
Further reading; Drug information, trough levels and immunogenicity.

Monday, May 16, 2016

Biosimilars: Similar but not identical. Contract manufacturing ELISA kits for your biosimilar (PK&ADA) and kits recently developed for CT-P13

Similar but not identical

A biosimilar is highly similar to, but not exactly the same as the existing, FDA-approved biologic, called the “reference” drug . People are familiar with generic versions of brand-name drugs, but biosimilars are not generic drugs. Generic versions of brand-name drugs are exact copies of chemically synthesized medicines. Biosimilars are not-quite-perfect copies of biologics – drugs derived from living cells that are impossible to replicate exactly.
Development of biosimilar proteins will present drug developers with a variety of challenges, both in manufacturing and in the clinic. Immunogenicity will prove to be one of the biggest challenges. Proper design and validation of an assay to detect anti-drug antibodies and accurate interpretation of sample analysis results will prove integral to developing a biosimilar protein.

Taking the above and below-mentioned into account Matriks Biotek® produced SHIKARI® Q-REMS and S-AIR as the  first ELISA kits for the biosimilar of Infliximab, CT-P13 (Remsima®)and now also offers to develop custom ELISA kits for your biosimilars or biologics on contract manufacturing basis depending on the molecule provided. Matriks Biotek® is the first company to commercialize SHIKARI® ELISA kits for biological drug testing to measure trough levels and anti drug antibodies (immunogenicity) since 2008. 

Comparisons for measuring especially ADA and trough levels are made by the tests developed for reference drug not for biosimilar which may differ on the test results. On the contrary we do not know the results when comparisons done by the ELISA kit developed for the biosimilar drug, it can present low immunogenicity as well.
The immunogenicity caused by the production of anti-drug antibodies (ADA) is also an important factor, and in many cases, an effect that prompts discontinuation of treatment. After long periods of treatment, ADA have been detected by the immunoenzymatic essay (ELISA). ADA may cause neutralization of the molecule, affecting PD and PK, making the treatment ineffective. 
The causes of immunogenicity can be chimeric biological drugs (e.g. infliximab), even humanized molecules (e.g. adalimumab) and fully humanized biological drugs (golimumab)—most the cases the residual immunogenicity resides in the CDR regions —glycosylation profiles, fermentation, purification, formulation (aggregate formation), administration mode (i.m., i.v. and s.c.), dosing, degradation products and contaminants.
The pharmaceutical formulation strategy is a critical step and needs to be accurate. The knowledge about physical and biological properties of the biological drug orientate the formulation process. Important components of protein formulations are pH, stabilizer, solubilizer, buffer, and tonicity modifier (bulking agent). The typical stability problems observed in protein pharmaceuticals are non-covalent aggregation, covalent aggregation, deamidation, cyclic imide, and cleavages. This process can affect directly the efficacy (e.g. immunogenicity) and safety (e.g. adverse events) of a biological drug.
The potential immunogenicity of a therapeutic protein can be influenced not only by the manufacturing processes mentioned above, but also by the type of disease, route of administration and dose. Biosimilar proteins will most likely not be identical to the innovator drug, and because of this, immunogenicity becomes a major concern when developing biosimilar proteins. The potential exists for serious clinical consequences due to anti-drug immune responses with all protein therapeutics. A safe immunogenicity profile of the innovator product does not indicate that the biosimilar protein will be safe. 
Proper immunogenicity assessment of biosimilars requires an in-depth understanding of the design of assays to detect anti-drug antibodies, implementation of proper validation procedures, and experience in analysis and interpretation of sample results. Use of an inappropriate anti-drug antibody assay format, improper validation testing or analysis of sample results may lead to misinterpretation of safety data. Anti-drug antibody assays must be properly designed and developed if they are to perform as intended. Many platforms and assay formats are available, and, depending on the protein therapeutic and its target, not all are appropriate for use. There are many variables that must be considered when developing an assay to detect anti-drug antibodies. The design of the clinical trial (single vs. multiple dose study), anticipated therapeutic drug levels in the immunogenicity samples, therapeutic target of the drug, mechanism of action of drug, and patient disease state all need to be considered when developing immunogenicity assays. Once the appropriate assay has been developed, the assay must be properly validated. Validation must include statistical determinations of both screening and confirmatory cut points. The use of arbitrarily set cut points to determine sample reactivity is improper and scientifically invalid. In order to ensure that the assay performs as expected when analyzing study samples, validation testing must also include assessment of signal precision, from which the in-study acceptance criteria can be generated. During sample analysis, data must be properly analyzed to ensure that the assay is performing as intended and that samples are not being reported as false negatives.