Annals of Forensic Research and Analysis

New Approach in Forensic Analysis – Biomolecular Computing Based Analysis of Significant Forensic Biomarkers

Mini Review | Open Access | Volume 1 | Issue 1

  • 1. Department of Chemistry and Biomolecular Science, Clarkson University, USA
  • 2. Department of Chemistry, University at Albany, SUNY, USA
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Corresponding Authors
Jan Halámek, Department of Chemistry, University at Albany, USA

Novel biochemical assays based on biomolecular information processing realized in enzyme-biocatalytic systems were developed for forensic analysis of various biomarkers. On-site analyses of biomarkers for recognition of ethnic origin and gender of biological fluids were developed. Future perspectives for forensic analysis and scientific challenges are discussed in the paper.


•    Biomarkers
•    Enzymes
•    Logic gates
•    Biocomputing
•    Biocatalytic cascades


Katz E, Halámek J (2014) New Approach in Forensic Analysis – Biomolecular Computing Based Analysis of Significant Forensic Biomarkers. Ann Forensic Res Anal 1(1): 1002.


Biochemical assay of biological substances found on a crime scene is an important part of forensic investigation aiming at identification of crime victims and suspects. A biochemistry/ molecular biology-based subarea of forensic analysis, forensic serology, deals with the complex task of gathering information on type, age, origin or gender from biological fluids (blood, saliva, etc.) found on a crime scene [1-5]. Modern forensic serology [6-8] relies on two major methods: immunoassay [8] and DNA [9]/RNA [10] analysis. While simple primary steps in this investigation (e.g., identification of human blood vs. other colored spots) can be easily done in field conditions, more specific analytical steps require sophisticated laboratory equipment and highly qualified staff [6]. This introduces time-delay in the investigation procedure and increases the complexity of the investigation itself. Despite the modern trend of designing portable equipment for on-site forensic biochemical analysis [6,11], including microfluidic [12] and lab-on-a-chip [13] systems, there is an obvious lack of onsite detection/characterization technologies, analogous to pointof-care diagnostic approaches, which have become common in medical areas (e.g. diabetes management, pregnancy tests, etc.). A novel approach to the analysis of biological substances for simplifying recognition of biomarkers in forensic applications can be borrowed from recent advances in unconventional chemical computing [14], specifically, from recently pioneered biomolecular computing systems [15]. Such biocomputing systems based on proteins/enzymes [16,17], DNA [18], RNA [19], DNAzymes [20] and whole cells [21] can perform logic operations processing multiple biochemical input signals. Based on biomolecular systems various Boolean logic operations such as AND, OR, XOR, NOR, NAND, INHIB, XNOR, etc., were realized [15]. Sophisticated networks composed of several concatenated biomolecular logic gates performing complex logic operations were designed for unconventional computing applications [15]. Particularly rapid progress was recently achieved in the area of enzyme-based biocomputing systems allowing their assembling in the form of concatenated logic networks composed of many gates performing various Boolean logic operations and including non-Boolean elements, such as enzyme-based multiplexer and demultiplexer, amplifier, filter, etc., mimicking their electronic counterparts [16]. Although chemical information processing systems were originally considered exclusively for computational applications, [22,23] it has recently been realized that they have features suitable for analytical/bioanalytical use [24,25]. These systems can analyze several biochemical signals according to a “preprogrammed” algorithm and generate a binary “YES/ NO” answer without using a computer. Such chemical testing, analogous to a pregnancy test, might be convenient for certain end-user and point-of-care applications, being particularly useful for on-site forensic investigations. Networks with computational steps that involve only biochemical processes [15,16] are being investigated for new technological capabilities that include multi-input biosensors with new functionalities [24,25]. After developing biomedical applications (e.g., analysis of injurysignaling biomarkers) [26-28] and military/homeland security applications (e.g., analysis of explosives and chemical weapons) [29], the biocomputing approach was successfully implemented to forensic applications [30,31].

The important feature of the novel analytical approach is the rapid qualitative analysis in the binary form “YES/NO” in response to the presence/absence of various combinations of biomarkers. Notably, instead of a quantitative analysis of a single highly specific biomarker, the present approach is based on the logic processing (e.g., in the format of Boolean AND logic gate) of several less specific biomarkers with all information processing steps performed by biochemical reactions, Figure 1A.

One of the new applications based on biocatalytic cascades analyzing simultaneous presence of creatine kinase (CK) and lactate dehydrogenase (LDH) was developed aiming at the recognition of biofluids of different ethnic origin for forensic analysis, Figure 1B [30]. Knowing the difference in the concentrations of CK and LDH in the blood of healthy adults of two ethnical groups, Caucasian (CA) and African American (AA), and taking into account the human population distribution patterns, samples of different ethnic origin with various CK/ LDH concentrations were mimicked and analyzed. The analysis was performed using a multi-enzyme / multi-step biocatalytic cascade where the differences in both included enzymes resulted in an amplified difference in the final analytical response. The statistically established analytical results confirmed excellent probability to distinguish samples of different ethnic origin (CA vs. AA). The results obtained on the model solutions were confirmed by the analysis of real serum samples collected from human subjects of different ethnic origin.

Another biocatalytic assay analyzing simultaneous presence of creatine kinase (CK) and alanine transaminase (ALT) was developed aiming at the recognition of biofluids of different gender for forensic applications, Figure 1C [31]. Knowing the difference in the concentrations of CK and ALT enzymes in the blood of healthy adults of male and female groups we mimicked the samples of different gender with various CK/ALT concentrations. The analysis was performed using a multi-enzyme / multi-step biocatalytic cascade where the differences in both included enzymes resulted in an amplified difference in the final analytical response. The analysis performed in human serum solutions allowed discrimination of samples corresponding to male/ female groups. The robustness of the developed analysis allowed determination of the gender for serum solutions after their drying and ageing at least for 1 hour. Importantly for forensic application, reaction with a chromogenic reactant nitroblue tetrazolium allowed qualitative discrimination of the “male” and “female” samples with a naked eye.

Applying novel concepts from the area of unconventional computing (specifically from biomolecular computing) [15,16] to biosensing and bioanalytical assays [24,25] has resulted in the design of biomolecular systems logically processing several chemical signals and converting them to a single binary output in the format of YES/NO. Information processing in biomolecular systems does not require electronic computers and proceeds at the level of chemical reactions. The “program” for processing chemical inputs can be implemented in the composition of the biomolecular system and can include various logic operations applied in different combinations. The systems exemplified above demonstrated the simplest logic processing based on logic AND/OR gates, applied to biochemical input signals. However, many other logic operations integrated in various logic circuitries are possible with the use of different enzymes [16] and other biomolecules [15], to allow high-fidelity detection of various biomarkers. The resulting digital biosensors would thus benefit different important fields, being particularly useful in rapid onfield forensic analysis. An important challenge in developing this kind of the digital multi-signal biosensor system is obtaining a significant difference between the logic 1 and 0 output values (in other words, a well-defined YES/NO answer). One should remember that in case of forensic analysis the input signals (biomarkers) appear with small difference. In order to obtain significant difference in the output signals, thus being able to distinguish biological samples of different origin, the responsefunction in the information processing steps should be sigmoid rather than linear [32-34]. In other words, the system should demonstrate a non-linearity with a sharp transition between the 0 (NO) and 1 (YES) states. The first steps in this direction have been already done experimentally and analyzed theoretically [35]. However, extensive research effort aimed at designing chemical “filter”-systems similar to the electronic counterparts is needed. The thresholds separating the logic 0 and 1 values could be tailored for a specific application by following expected concentrations of biomarkers in the biological fluids. This will be an important step towards future on-site forensic analysis.


This work was supported by the NSF awards # CBET1066397.


1. Li R. Forensic Biology. CRC Press, Boca Raton, FL. 2008.

2. Shaler RC. Modern forensic biology. In: Saferstein R, editor. Forensic Science Handbook. Prentice Hall, Upper Saddle River. 2002; 529-546.

3. Kobilinsky L. Forensic Chemistry Handbook. , NJ: Wiley, Hoboken. 2011.

4. Sikirzhytski V, Sikirzhytskaya A, Lednev IK. Multidimensional Raman spectroscopic signatures as a tool for forensic identification of body fluid traces: a review. Appl Spectrosc. 2011; 65: 1223-1232.

5. Virkler K, Lednev IK. Raman spectroscopic signature of semen and its potential application to forensic body fluid identification. Forensic Sci Int. 2009; 193: 56-62.

6. Virkler K, Lednev IK. Analysis of body fluids for forensic purposes: from laboratory testing to non-destructive rapid confirmatory identification at a crime scene. Forensic Sci Int. 2009; 188: 1-17.

7. An JH, Shin KJ, Yang WI, Lee HY. Body fluid identification in forensics. BMB Rep. 2012; 45: 545-553.

8. Gaensslen RE. Sourcebook in Forensic Serology, Immunology, and Biochemistry. U.S. Department of Justice, Washington, DC. 1983.

9. Elkins KM. Forensic DNA Biology: A Laboratory Manual. Academic Press, Oxford, UK. 2012.

10. Bauer M. RNA in forensic science. Forensic Sci Int Genet. 2007; 1: 69- 74.

11. Eckenrode A, Bartick EG, Harvey S, Vucelick ME, Wright BW, Huff RA. Portable Raman spectroscopy systems for field analysis. Forensic Sci. Commun. 2001; 4.

12. Horsman KM, Bienvenue JM, Blasier KR, Landers JP. Forensic DNA analysis on microfluidic devices: a review. J Forensic Sci. 2007; 52: 784-799.

13. Liu P, Yeung SH, Crenshaw KA, Crouse CA, Scherer JR, Mathies RA. Real-time forensic DNA analysis at a crime scene using a portable microchip analyzer. Forensic Sci Int Genet. 2008; 2: 301-309.

14. Katz E. Molecular and Supramolecular Information Processing: From Molecular Switches to Logic Systems. Wiley-VCH, Weinheim, Germany. 2012.

15. Katz E. Biomolecular Information Processing - From Logic Systems to Smart Sensors and Actuators. Wiley-VCH, Weinheim, Germany. 2012.

16. Katz E, Privman V. Enzyme-based logic systems for information processing. Chem Soc Rev. 2010; 39: 1835-1857.

17. Unger R, Moult J. Towards computing with proteins. Proteins. 2006; 63: 53-64.

18. Ezziane Z. DNA computing: Applications and challenges. Nanotechnology 2006; 17: R27-R39.

19. Win MN, Smolke CD. Higher-order cellular information processing with synthetic RNA devices. Science. 2008; 322: 456-460.

20. Stojanovic MN, Mitchell TE, Stefanovic D. Deoxyribozyme-based logic gates. J Am Chem Soc. 2002; 124: 3555-3561.

21. Li Z, Rosenbaum MA, Venkataraman A, Tam TK, Katz E, Angenent LT. Bacteria-based AND logic gate: a decision-making and self-powered biosensor. Chem Commun (Camb). 2011; 47: 3060-3062.

22. Benenson Y. Biocomputers: from test tubes to live cells. Mol Biosyst. 2009; 5: 675-685.

23. Stojanovic MN. Some experiments and directions in molecular computing and robotics. Isr. J. Chem. 2011; 51: 99-105.

24. Wang J, Katz E. Digital biosensors with built-in logic for biomedical applications-biosensors based on a biocomputing concept. Anal Bioanal Chem. 2010; 398: 1591-1603.

25. Katz E, Wang J, Privman M, Halámek J. Multianalyte digital enzyme biosensors with built-in Boolean logic. Anal Chem. 2012; 84: 5463- 5469.

26. Halámek J, Windmiller JR, Zhou J, Chuang M-C, Santhosh P, Strack G, et al. Multiplexing of injury codes for the parallel operation of enzyme logic gates. Analyst. 2010; 135: 2249-2259.

27. Halámek J, Bocharova V, Chinnapareddy S, Windmiller JR, Strack G, Chuang MC, et al. Multi-enzyme logic network architectures for assessing injuries: digital processing of biomarkers. Mol Biosyst. 2010; 6: 2554-2560.

28. Zhou J, Halámek J, Bocharova V, Wang J, Katz E. Bio-logic analysis of injury biomarker patterns in human serum samples. Talanta. 2011; 83: 955-959.

29. Chuang MC, Windmiller JR, Santhosh P, Valdés Ramírez G, Katz E, Wang J. High-fidelity determination of security threats via a Boolean biocatalytic cascade. Chem Commun (Camb). 2011; 47: 3087-3089.

30. Kramer F, Halámková L, Poghossian A, Schöning MJ, Katz E, Halámek J. Biocatalytic analysis of biomarkers for forensic identification of ethnicity between Caucasian and African American groups. Analyst. 2013; 138: 6251-6257.

31. Bakshi S, Halámková L, Halámek J, Katz E. Biocatalytic analysis of biomarkers for forensic identification of gender. Analyst. 2014; 139: 559-563.

32. Bakshi S, Zavalov O, Halámek J, Privman V, Katz E. Modularity of biochemical filtering for inducing sigmoid response in both inputs in an enzymatic AND gate. J Phys Chem B. 2013; 117: 9857-9865.

33. Privman V, Fratto BE, Zavalov O, Halámek J, Katz E. Enzymatic AND logic gate with sigmoid response induced by photochemically controlled oxidation of the output. J Phys Chem B. 2013; 117: 7559- 7568.

34. Zavalov O, Bocharova V, Privman V, Katz E. Enzyme-based logic: OR gate with double-sigmoid filter response. J Phys Chem B. 2012; 116: 9683-9689.

35. Privman V. Control of noise in chemical and biochemical information processing. Isr. J. Chem. 2011; 51: 118-131

Katz E, Halámek J (2014) New Approach in Forensic Analysis – Biomolecular Computing Based Analysis of Significant Forensic Biomarkers. Ann Forensic Res Anal 1(1): 1002.

Received : 24 Feb 2014
Accepted : 28 Feb 2014
Published : 04 Mar 2014
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