Research Activities I. Iron metabolism macromolecular trafficking and protein multifunctionality Though iron is an essential micronutrient for all life it is a double edged sword, consequently its homeostasis is a tightly regulated process to maintain the delicate balance between iron nutrition and toxicity. Practically all of extracellular iron is chelated to the iron carrier proteins transferrin (an abundant serum iron transport protein) and lactoferrin (present in other extracellular fluids) Cells of the immune system are known to acquire iron for metabolic use from transferrin via endocytosis of transferrin- iron complexes. Until recently two receptors had been identified for transferrin uptake(Transferrin receptor 1 & Transferrin receptor 2) however the existence of additional receptors was long suspected. Our laboratory identified a novel receptor for this vital process. Interestingly it has been identified as Glyceraldehyde-3-phosphate dehydrogenase (GAPDH). This enzyme has long been known as one of the key enzymes of the glycolytic pathway. It is abundantly present in all cells and has been classified as a housekeeping gene. Over the last few years many new functions have attributed to GAPDH, and it is now well recognized as a moonlighting (multifunctional) protein. My group has characterized this novel iron carrier protein receptor in detail and have found it to exhibit higher order multifunctionality. We have demonstrated that apart from functioning as a cell membrane surface receptor cells also actively secrete GAPDH upon iron starvation. This secreted GAPDH (sGAPDH) efficiently traffics transferrin into cells in an autocrine and paracrine manner. Other investigations from our laboratory have also demonstrated GAPDH to function as a dual receptor for lactoferrin. Iron is an essential element for vital metabolic processes but is toxic in excess, consequently its homeostasis is a tightly regulated process. Though the mechanisms of iron import into cells are now relatively well characterized, iron export from cells still remains poorly understood. Till date ferroportin is the only known mammalian iron exporter and is expressed in duodenal enterocytes, macrophages and hepatocytes. In addition to understanding the role of GAPDH in iron acquisition by cells our group has also uncovered an additional dimension to the multifunctionality of GAPDH where, to maintain cellular homeostasis, the same protein behaves in a converse manner under two opposing conditions of cellular iron status. Using cell types that play a key role in maintaining iron homeostasis (cells of reticuloendothelial system, hepatocytes and enterocytes) we have provided evidence that cells continue to enhance their recruitment of GAPDH to the membrane surface when exposed to iron overload. This GAPDH is of a different isoform than that recruited upon iron starvation and does not bind Holo Transferrin (the iron replete form of transferrin), instead it interacts with Apo Transferrin (the iron free form of transferrin) with high affinity and facilitates the export of iron from cells in a process involving close association with ferroportin. These findings have been validated in vivo utilizing rodent models of iron overload. As iron is vital for the replication and survival of invading pathogens during infection, we are also interested in understanding the mechanisms involved in this process using suitable infection models. To survive within the host cells successful intracellular pathogens employ several strategies to acquire host iron. Mycobacterium tuberculosis is one of the most successful intracellular pathogens. It is well known that the survival of M. tuberculosis within the macrophage depends critically on the availability of iron. Intracellular M.Tb utilizes several mechanisms to acquire iron by hijacking the host trafficking pathways. Our laboratory is studying these aspects by infecting macrophages with fluorescent fusion protein labelled M.Tb and evaluating mammalian GAPDH trafficking in these infected cells using, confocal microscopy, biochemical methods and electron microscopy. Significance of our work Iron deficiency is one of the most common and intractable nutritional problems affecting over 2 billion people in the world today. This has important consequences for human health (especially women) and child development. A compromised physical work capacity leads to overall economic and societal losses, especially among developing nations. The major co-morbidities associated with this condition are: compromised immunity and infectious dis¬eases like Tuberculosis. Paradoxically, iron supplementation worsens the course of infection in these patients. Our work has brought to light an evolutionarily ancient but hitherto unknown dimension of the mechanisms by which organisms regulate their iron acquisition. It also throws light on the conflict between, mammalian host and resident pathogen in the struggle to acquire this vital resource for oneself while denying its access to the other. The results of our work can now be exploited to develop effective mechanisms to deliver nutritionally vital iron while at the same time disrupting the pathogens iron uptake mechanisms thereby obtaining the double benefits of nutritional amelioration along with control of infection. II. Development of optimization strategies for quantitative localization of antigens in biological samples by electron microscopy Choice of reagents and sample processing protocols is crucial for the efficient detection and quantification of target antigen in cell & tissue by immunogold labeling electron microscopy. This is especially so when sample and/or reagents are available in limited amounts. Utilizing a recombinant intracellular protein antigen as model system we have developed ELISA based methods for quantitative monitoring of antigen loss in response to various processing steps in immuno electron microscopy. This work has also been extended to evaluate ‘background signals’ that are encountered during immunolabeling thus aiding in the selection of optimal processing steps and suitable blocking agents for quantitative ultrastructural localization of protein antigens. A prior knowledge, regarding the presence of target antigens and the ability of reagents to detect them, is extremely useful in carrying out ultrastructural localization of any antigen in cells and tissues. For achieving this goal we have developed a TEM based solid phase assay method that utilizes ultra low volume of reagents. Suspensions of cells and organelles, which have to be subjected to investigation by electron microscopy, present some unique challenges in terms of handling during sample processing. In my laboratory a versatile method, based upon immobilization on charged organic membranes, which allows for such material to be subjected to a wide variety of microscopic (light & EM) analysis has been developed. Another area of development in our laboratory has been to carry out correlative studies of macromolecular quantitation in biological samples using a multi approach strategy involving TEM, AFM and live cell imaging via confocal microscopy. Interaction between target molecules is cross-correlated via epitope based immunolabelling EM, force spectroscopy and FRET analysis. These studies have found application in studying the quantitative distribution/trafficking of various proteins in different cellular compartments in cells as part of our own work as well as that undertaken in collaboration with other groups. III. Development of immunobiosensors for detection of organophosporous pesticides The objective of this work carried out in colloboration with two other groups of our institute has been to develop a specific and sensitive methodology for monitoring toxic molecules (pesticides) from soil, water and agro-products using an immunobiosensor technique. Specific antibodies were raised and immobilized on a sensor surface. After high-resolution characterization of the bio-interface, using Atomic Force & Scanning Tunneling microscopy the target toxins were quantitated using piezoelectric microgravimetric assay as well as surface plasmon resonance techniques. The forces of interaction between immobilized molecules and antibody was studied by force spectroscopy. Though this work was primarirly targeted towards development of commercial technology and has yeilded several patents, the basic research that had to be undertaken for this task has also resulted in numerous publications. IV. Intracellular trafficking in infected macrophages Phagocytic cells like macrophages take up invading microorganisms and target them to the lysosomes for destruction. However intracellular pathogens like salmonella, etc. bypass the lysosomal transport and continue to survive in macrophages. In collaboration with a group at the National Institute of Immunology, New Delhi and another at IMTECH we have carried out macromolecular trafficking studies utilizing immunolabeling electron microscopy to understand the fundamental mechanism of how intracellular bacteria bypass lysosomal targeting. V. Electron Microscopy of protein complexes In collaboration with several research groups we have carried out high resolution analytical analysis of several protein complexes including virus like proteins (VLPs), membrane pore forming proteins and amyloid aggregates. Significant recognition: Awards, fellowships, international funding of distinction, technologies transferred/licensed etc.: Senior Research Associateship of the National Research Council (NRC) USA, 2003-2004. Selected list of Publications and Patents: Publications representative of above research topics: 1.Moonlighting glycolytic protein glyceraldehyde-3-phosphate dehydrogenase (GAPDH): An evolutionarily conserved plasminogen receptor on mammalian cells. Anoop Singh Chauhan, Manoj Kumar, Surbhi Chaudhary, Anil Patidar, Asmita Dhiman, Navdeep Sheokand, Himanshu Malhotra, Chaaya I. Raje and Manoj Raje. The FASEB Journal, Manuscript in Press: DOI10.1096/fj.201600982R Online Publication Date Issue 2017-06-01 Volume 31 Issue No.6. 2. Mycobacterium tuberculosis Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) functions as a receptor for human lactoferrin Himanshu Malhotra, Anil Patidar, Vishant Boradia, Rajender Kumar, Ajay Kumar, Rakesh Dilip Nimbalkar, Zahid Gani, Rajbeer Kaur, Prabha Garg, Manoj Raje and Chaaya Iyengar Raje Frontiers in Cellular and Infection Microbiology, (2017) manuscript in press. 3. Reverse overshot water-wheel retroendocytosis of Apo Transferrin extrudes cellular iron. Navdeep Sheokand, Himanshu Malhotra, Anoop Singh Chauhan, Manoj Kumar, Surbhi Chaudhary, Anil Patidar, Vishant Mahendra Boradia, Chaaya Iyengar Raje, and Manoj Raje Journal of Cell Science; 129, 843-853 doi: 10.1242/jcs.180356, (2016). 4.Exosomes: tunable nano vehicles for macromolecular delivery of transferrin and lactoferrin to specific int racellular compartment. Himanshu Malhotra, Navdeep Sheokand, Santosh Kumar, Anoop S. Chauhan, Manoj Kumar, Priyanka Jakhar, Vishant M. Boradia, Chaaya I. Raje and Manoj Raje Journal of Biomedical Nanotechnology; 2016, 12, 1101-1114. 5. Purification and Characterization of Glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) from Pea Seeds. Zahid Gani, Vishant M Boradia, Raghu R Janaki, Prashant M Suryavanshi, Pravinkumar H Patil, Santosh Kumar, Ranvir Singh, Manoj Raje and Chaaya Iyengar Raje. Protein Expression and Purification, 2016, Jul 4;127: 22-27.doi:10.1016/j.pep.2016.06.014 6. Mycobacterium tuberculosis H37Ra: a surrogate for the expression of conserved, multimeric proteins of M.tb H37Rv. Vishant Mahendra Boradia, Pravinkumar Patil, Anushri Agnihotri, Ajay Kumar, Kalpesh Kumar Rajwadi, Ankit Sahu, Naveen Bhagath, Navdeep Sheokand, Manoj Kumar, Himanshu Malhotra, Rachita Patkar, Navi Hasan, Manoj Raje and Chaaya Iyengar Raje Microbial Cell Factories; 2016, 15:140 DOI 10.1186/s12934-016-0537-0 7. Secreted multifunctional Glyceraldehyde-3-phosphate dehydrogenase sequesters lactoferrin and iron into cells via a non-canonical pathway. Anoop S. Chauhan, Pooja Rawat, Himanshu Malhotra, Navdeep Sheokand, Manoj Kumar, Anil Patidar, Surbhi Chaudhary, Priyanka Jakhar, Chaaya I. Raje and Manoj Raje Scientific Reports 5, 18465; doi:10.1038/srep18465 (2015). 8. Functional characterization of monomeric GTPase Rab1 in the secretory pathway of Leishmania Surbhi Bahl, Smriti Parashar, Himanshu Malhotra, Manoj Raje, and Amitabha Mukhopadhyay The Journal of Biological Chemistry; 2015, 290(5): 29993–30005. 9. Moonlighting cell surface GAPDH recruits Apo Transferrin to effect iron egress from mammalian cells Navdeep Sheokand, Himanshu Malhotra, Santosh Kumar , Vikas Ajit Tillu, Anoop Singh Chauhan, Chaaya Iyengar Raje and Manoj Raje Journal of Cell Science; 2014,127, 4279–4291 doi:10.1242/jcs.154005 10. Mycobacterium tuberculosis acquires iron by cell surface sequestration and internalization of human holo-transferrin Vishant Mahendra Boradia, Himanshu Malhotra, Janak Shrikant Thakkar, Vikas Ajit Tillu, Bhavana Vuppala, Pravinkumar Patil, Navdeep Sheokand, Prerna Sharma, Anoop Singh Chauhan, Manoj Raje and Chaaya Iyengar Raje Nature Communications; 2014, 5:4730 doi: 10.1038/ncomms5730. 11. Protein moonlighting in iron metabolism: glyceraldehyde-3-phosphate dehydrogenase (GAPDH) Vishant Mahendra Boradia, Manoj Raje, Chaaya Iyengar Raje Biochemical Society Transactions; 2014, 42(6): 1796-1801. 12. Calcium binding to Beta-2-microglobulin at physiological ph drives the occurrence of conformational changes which cause the protein to precipitate into amorphous forms that subsequently transform into amyloid aggregates. Kumar S, Sharma P, Arora K, Raje M, Guptasarma P. PLoS One. 2014 Apr 22;9(4):e95725. doi: 10.1371/journal.pone.0095725. eCollection 2014. 13. Secreted glyceraldehye-3-phosphate dehydrogenase is a multifunctional autocrine transferrin receptor for cellular iron acquisition Navdeep Sheokand, Santosh Kumar, Himanshu Malhotra, Vikas Tillu, Chaaya Iyengar Raje, and Manoj Raje Biochimica et Biophysica Acta (BBA)-General Subjects; 2013, 1830: 3816–3827 . 14. Clathrin-mediated hemoglobin endocytosis is essential for survival of Leishmania Shruti Agarwal, Ruchir Rastogi, Deepika Gupta, Nitin Patel, Manoj Raje and Amitabha Mukhopadhyay Biochimica et Biophysica Acta (BBA)-Molecular Cell Research; 2013, 1833: 1065–1077. 15. The multifunctional glycolytic protein glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a novel macrophage lactoferrin receptor. Pooja Rawat, Santosh Kumar, Navdeep Sheokand, Chaaya Iyengar Raje and Manoj Raje. Biochemistry and Cell Biology; 2012, 90(3): 329–338. 16. Characterization of Glyceraldehyde-3-phosphate dehydrogenase as a novel transferrin receptor. Santosh Kumar, Navdeep Sheokand, Mayur Anant Mhadeshwar, Chaaya Iyengar Raje and Manoj Raje. The International Journal of Biochemistry & Cell Biology; 2012, 44:189– 199 17. IL-6 inhibits IFN-γ induced autophagy in Mycobacterium tuberculosis H37Rv infected macrophages. Rajesh Kumar Dutta, Mahesh Kathania, Manoj Raje, Sekhar Majumdar The International Journal of Biochemistry & Cell Biology; 2012, 44: 942–954. 18. Exploitation of marine bacteria for production of gold nanoparticles Nishat Sharma, Anil K Pinnaka, Manoj Raje, Ashish FNU, Mani S Bhattacharyya and Anirban R Choudhury Microbial Cell Factories; 2012, 11:86 doi:10.1186/1475-2859-11-86 19. Bfl-1/A1 acts as a negative regulator of autophagy in mycobacteria infected macrophages. Kathania M, Raje CI, Raje M, Dutta RK, Majumdar S. The International Journal of Biochemistry & Cell Biology; 2011, 43(4):573-85. 20. Responses of Mycobacterium tuberculosis hemoglobin promoters to in vitro and in vivo growth conditions. Pawaria S, Lama A, Raje M, Dikshit KL. Applied and Environmental Microbiology; 2008, 3512-3522. 21. Inhibition of bfl-1/A1 by siRNA inhibits mycobacterial growth in THP-1 cells by enhancing phagosomal acidification. Dhiman R, Kathania M, Raje M, Majumdar S. Biochim Biophys Acta.; 2008,1780:733-742. 22. Coalescence of spherical beads of retro-HSP12.6 into linear and ring-shaped amyloid nanofibers. Anshuman Shukla, Manoj Raje and Purnananda Guptasarma. Biochemistry (Moscow); 2008, 7396):681-685. 23. The macrophage cell surface Glyceraldehyde-3-phosphate dehydrogenase is a novel Transferrin receptor. Chaaya Iyengar Raje, Santosh Kumar, Arti Harle, Jagpreet Singh Nanda and Manoj Raje The Journal of Biological Chemistry; 2007, 282(5): 3252–3261. 24. Nitric oxide induces apoptosis in cutaneous T cell lymphoma (HuT-78) by downregulating constitutive NF-kappaB. Loovena Rishi, Rohan Dhiman, Manoj Raje M and Sekhar Majumdar Biochim Biophys Acta.; 2007,1770(8):1230-9. 25. Differential expression of NF-kappaB in mycobacteria infected THP-1 affects apoptosis. Rohan Dhiman, Manoj Raje and Sekhar Majumdar Biochim Biophys Acta.; 2007, 1770(4):649-58. 26. Immunochromatographic Dipstick Assay Format Using Gold Nanoparticles Labeled Protein-Hapten Conjugate for the Detection of Atrazine. Jasdeep Kaur, K .Vikas Singh, Robin Boro, K.R.Thampi, Manoj Raje, Grish C. Varshney and C. Raman Suri Environ. Sci. Technol.; 2007, 41: 5028-5036 27. Charged nylon membrane substrate for convenient and versatile high resolution microscopic analysis of E.coli & mammalian cells in suspension culture Manoj Raje, Rohan Dhiman, Sekhar Majumdar, Taposh Dass, Kanak L. Dikshit and Ramandeep Kaur Cytotechnology; 2006, 51:111-117 28. Atomic Force Spectroscopy based study of antibody pesticide interactions for characterization of immunosensor surface. Kaur J, Singh KV, Schmid AH, Varshney GC, Suri CR and Manoj Raje Biosensors and Bioelectronics; 2004, 20(2):284-93. 29. Strategies for direct attachment of hapten to polystyrene support for applications in enzyme-linked immunosorbent assay (ELISA). J. Kaur, K V Singh, Manoj Raje, G. C. Varshney and C R.Suri. Anal. Chim. Acta.; 2004, 506, 133-135. 30. Synthesis and characterization of Hapten-Protein conjugates for antibody production against small molecules. K.V. Singh J Kaur, G.C Varshney, Manoj Raje and C.R. Suri Bioconjungate Chem.; 2004, 15, 168-173. 31. A Solid Phase method for evaluation of gold conjugate used in quantitative detection of antigen by immunogold labeling electron microscopy Ramandeep Kaur and Manoj Raje Journal of Immunological Methods; 2003, 279(1-2):33-40. 32. A backbone-reversed form of an all-beta alpha-crystallin domain from a small heat-shock protein (retro-sHSP12.6) folds and assembles into structured multimers Anshuman Shukla, Manoj Raje, and Purnananda Guptasarma The Journal of Biological Chemistry; 2003, 278(29): 26505-26510. 33. An ELISA based approach to optimize elution conditions for screening antibodies against hapten K V Singh, Jasdeep Kaur, Manoj Raje, Grish C Varshney & C Raman Suri Anal Bioanal Chem.; 2003, 377: 220-224. 34. A backbone-reversed all-beta polypeptide (retro-CspA) folds and assembles into amyloid nanofibres. Shukla A, Raje Manoj and Guptasarma P. Protein Eng.; 2003, 16(12): 875-9. 35. Pre- screening for antigen detectability in cells: A TEM based solid phase digital immunogold detection method utilizing ultra low volumes of reagents Ramandeep Kaur, Kanak L, Dikshit and Manoj Raje The Journal of Microscopy; 2002, 208 (2), 100-107. 36. Optimization of Immunogold Labeling TEM: An ELISA-based method for evaluation of Blocking Agents for Quantitative Detection of Antigen Ramandeep Kaur, Kanak L. Dikshit and Manoj Raje The Journal of Histochemistry & Cytochemistry; 2002, 50: 863-874. 37. Immunosensor for pesticide analysis: Antibody production and sensor development C. Raman Suri, Manoj Raje & Grish C. Varshney Critical Reviews In Biotechnology; 2002, 22(1): 15-32. 38. Sandwich microgravimetric immunoassay: Sensitive and specific detection of low molecular weight analytes using piezoelectric quartz crystal. Satish Saha, Manoj Raje and C.Raman Suri. Biotechnology Letters; 2002, 24: 711-716. 39. Evidence that an eukaryotic-type serine/threonine protein kinase from Mycobacterium tuberculosis regulates morphological changes associated with cell division. Chaba, R., Manoj Raje & Chakraborti, P. K. European Journal of Biochemistry; 2002, 269: 1078-1085. 40. Evidence that glycoprotein 96 (b2), a stress protein, functions as a th2-specific costimulatory molecule. Banerjee PP, Vinay DS, Mathew A, Raje M, Parekh V, Prasad DV, Kumar A, Mitra D, Mishra GC. J Immunol; 2002,169(7):3507-18. 41. Recombinant dengue virus type 2 envelope/hepatitis B surface antigen hybrid protein expressed in Pichia pastoris can function as a bivalent immunogen. Bisht H, Chugh D, Raje M, Swaminathan S, Khanna N. J Biotechnol; 2002, 99(2):97-110. 42. Vitreoscilla Hemoglobin: Intracellular Localization and Binding to Membranes. Ramandeep, Kwang Woo Hwang, Manoj Raje, Kyung¬Jin Kim, Benjamin C. Stark, Kanak L. Dikshit and Dale A. Webster. The Journal of Biological Chemistry; 2001, 276: 24781-24789. 43. SopE acts as a Rab5 specific nucleotide exchange factor and recruits nonprenylated Rab5 on Salmonella containing phagosomes to promote fusion with early endosome. Konark Mukherjee, Parashuraman Seetharaman, Manoj Raje, and Amitabha Mukhopadhyay The Journal of Biological Chemistry; 2001, 276: 23607-23615. 44. Optimization of Immunogold Labeling TEM: An ELISA based Method for Rapid and Convenient Simulation of Processing Conditions for Quantitative Detection of Antigen. Ramandeep, Kanak L. Dikshit and Manoj Raje The Journal of Histochemistry & Cytochemistry; 2001, 49(3): 355-367. 45. Live Salmonella Modulate Expression of Rab Proteins to persist in a Specialized Compartment and Escape Transport to Lysosomes. Shehla Hashim, Konark Mukherjee, Manoj Raje, Sandip K. Basu and Amitabha Mukhopadhyay. The Journal of Biological Chemistry; 2000, 275(21): 16281-8. 46. Live Salmonella Recruits N-Ethylmaleimide-sensitive Fusion Protein on Phagosomal Membrane and Promotes Fusion with Early Endosome. Konark Mukherjee, Shadab A. Siddiqi, Shehla Hashim, Manoj Raje, Sandip K. Basu and Amitabha Mukhopadhyay. The Journal of Cell Biology; 2000, 148(4): 741-753. 47. Hemoglobin Endocytosis in Leishmania Is Mediated through a 46-KDa protein Located in the Flagellar Pocket. Shantanu Sengupta, Jalaj Tripathi, Ruchi Tandon, Manoj Raje, Rajendra P. Roy, Sandip K. Basu and Amitabha Mukhopadhyay. The Journal of Biological Chemistry; 1999, 274(5): 2758-2765. 48. Localization of DNAK & GroEL in Vibro Cholerae. Jeevan Jyot, Jitendra Kumar Gautam, Manoj Raje and Amit Ghosh. FEMS Microbiology Lett.; 1999, 172 : 165-171. 49. Routes of Transmission in the Hepatitis E Epidemic of Saharanpur. V. Singh, V. Singh, M. Raje, C.K. Nain, K. Singh. Tropical Gastroenterology; 1998, 19 : 107-109. 50. The Role of N-terminal in the kinetics and assembly of a-hemolysin of Staphylococcus aureus. Vandana S., Manoj Raje and Krishna Sastry, M. The Journal of Biological Chemistry; 1997, 272: 24858-24863. 51. DNA Binding Protein Of Mycobacteria And Human Response. H.K. Prasad, S. Prabhakar, P.S. Annapurna, Shamsuzzaman, Surinder Kaur, N.K. Jain, A.B. Dey, Najmul Islam and Manoj Raje. Indian Journal of Clinical Biochemistry; 1997,12 (Suppl.): 86-88. 52. Recognition of the Parasite infected cell surface determinants by homologous antiserum raised against infected cell membranes. Amit Choudhury, Apollina Goel, Manoj Raje, Harpreet Vohra and Girish C. Varshney. Parasitology Research; 1997, 83:746-754. 53. Characterization of Novel Costimulatory Molecules: A protein of 38-42kD from B-cell surface is concerned with T-cell activation and differentiation. Dass S. Vinay, Manoj Raje, Rakesh Verma, and Gyan C. Mishra. The Journal of Biological Chemistry; 1996, 270: 1-14. 54. Characterization of a Novel Co-stimulatory molecule: a 155-160 kD B cell surface protein provides accessory help to CD4+ T cells to proliferate and differentiate. Dass S. Vinay, Manoj Raje, and Gyan C. Mishra. Molecular Immunology; 1996, 33: 1-14. 55. Attempts to characterize the mechanisms involved in the growth inhibition of Mycobacterium microti in interferon-g or tumor necrosis factor-a activated J774A.1 cells. Reena Gupta, Nisha Dogra, Manoj Raje and Shekhar Majumdar. FEMS Microbiology Letters; 1996,140: 171-178. 56. Identification of a novel 30 kDA antigen of M. Tuberculosis (MTB) Prasad, H.K. Annapurna, P.S. Zzaman, S, Najmul, L and Manoj Raje Journal of Cellular Biochemistry, 1995, Supplement 19B. 57. Determination of immunoglobulin M concentration by piezoelectric crystal immunobiosensor coated with protamine. C. Raman Suri, Manoj Raje & Gyan C. Mishra. Biosensors & Bioelectronics; 1994, 9: 535-542. Invited Chapters 1. Boradia VM, Raje M and Raje CI. Cell Surface Mycobacterium tuberculosis GAPDH functions as a transferrin receptor. Invited book chapter in Moonlighting Proteins: Novel Virulence Factors in Bacterial Infection. Wiley Blackwell, 2017, Editor. Brian Henderson. Chapter 11, pp: 205-224. 2. E-Pathshala-Biophysics for Post Graduate students, Ministry of Human Resources Development, Govt. of India E-book initiative,2015. Module on Light Microscopy and Fluorescence Microscopy 3. DNA Binding Protein HLPMt of Mycobacterium Tuberculosis A Target of Human Immune Response.Savita P, K.Raha, P.S.Annapurna, S.Kaur, Najmul Islam, S.Zzaman, Manoj Raje, A.B.Dey, N.K.Jain, V.Kothekar, J.S.Tyagi and H.K.Prasad. Invited chapter in Immunomodulation, ed. S.N. Upadhyay, Narosa Publishing House, New Delhi, 1999. 4. Autoradiography Manoj Raje and Gyan C.Mishra. Invited chapter in Encyclopedia of Immunology, Second Edition, ed. Peter J. Delves and Ivan M. Roitt. Academic Press Ltd. pp: 296-300, 1998. Patents Total 10 patents (8 International + 2 Indian) Selected Patents 1. Microporous filtration based dot immunoassay device for method for screening of analytes and method of use. US Patent. 2. An efficient and improved method for screening of analytes by microporous filtration based dot immunoassay kit. European Patent. 3. A dipstic based immunoassay for pesticide monitoring. Australian Patent.

Cell Biology, Biophysics, Confocal & Electron Microscop