Polyplex |Summer Training Report |

REVIEW OF LITERATURE
Polyplex Evolution: Understanding Biology,Optimizing Performance

Polyethylenimine (PEI) is a gold standard polycationic transfectant. However, the highly efficient transfecting
activity of PEI and many of its derivatives is accompanied by serious cytotoxic complications and safety concerns at
innate immune levels, which impedes the development of therapeutic polycationic nucleic acid carriers in general
and their clinical applications. In recent years, the dilemma between transfection efficacy and adverse PEI activities
has been addressed from in-depth investigations of cellular processes during transfection and elucidation of
molecular mechanisms of PEI-mediated toxicity and translation of these integrated events to chemical
engineering of novel PEI derivatives with an improved benefit-to-risk ratio. This review addresses these
perspectives and discusses molecular events pertaining to dynamic and multifaceted PEI-mediated cytotoxicity,
including membrane destabilization, mitochondrial dysfunction, and perturbations
of glycolytic flux and redox homeostasis as well as chemical strategies for the generation of better tolerated
polycations. We further examine the effect of PEI and its derivatives on
complement activation and interaction with Toll-like receptors. These perspectives are intended to lay the
foundation for an improved understanding of interlinked mechanisms controlling transfection and toxicity and
their translation for improved engineering of polycation-based transfectants.

Polyethylenimine (PEI) is among the most intensively investigated polycations for nucleic acid delivery. Because of
its highly efficient transfection capabilities, PEI has served as a gold standard transfectant polymer.13 PEI is
structured from repeating units of two aliphatic carbon groups and amino nitrogen and is commercially available in
both linear and branched morphologies, with the molecular weight
ranging between 200 Da and 1,500 kDa.13–18 Shortly after the discovery of branched PEI as a promising
transfecting agent, linear PEI, produced by hydrolysis of poly(2-ethyl-2-oxazoline), was identified as a
derivative with more favorable properties.19–21 The branched PEI contains primary, secondary, and tertiary
amines, whereas the linear form has only secondary amines. The protonable amine groups pro-
vide PEIs with the unique features of high cationic charge density and buffering capacity at extracellular and endo-
lysosomal pH levels.
Polyplex |Summer Training Report |

Conclusions

Since the first discovery of PEI as a transfecting agent, both the bio-logical assessment techniques for
the identification of subcellular impairment and the chemical engineering of new synthetic materials
have dramatically advanced. The scientific knowledge and methodologies from both perspectives can
contribute to the development of next-generation nucleic acid carriers with suitable safety profiles
that meet the demands on potential future therapeutics. As a result,new polymers have to be
investigated in comprehensive assessments to identify effects on the cellular metabolic situation and
detect subtle toxicity. For clinical applications, several other aspects beyond in vitrocytotoxicity also
have to be considered to ensure clinical safety: serum interaction, hemolysis, tissue distribution,
complement activation,metabolism, excretion, and more affect the suitability of polymers and
polyplexes for in vivo usage. Also, here, the identification of underlying mechanisms and establishment
of structure-activity relationships is key to the rational design and optimization of materials with
intended properties. Finally, a recent study demonstrated that
sub-50-nm PEI polyplexes can reach the nuclear membrane and bind exclusively to the nuclear pore
complex without causing disruption of the nuclear envelope.161 However, it is still necessary to eluci-
date whether PEI and DNA dissociate before transcription of DNA or whether transcriptional complexes
may bind to DNA while DNA remains bound to PEI and whether PEI in the nucleus can trigger
cell death.

AUTHOR CONTRIBUTIONS
All authors analyzed and discussed the literature. All authors contrib-
uted to the writing of the paper and revised the manuscript.

CONFLICTS OF INTEREST
The authors declare no competing financial interest.

ACKNOWLEDGMENTS
A.H. and J.B. acknowledge financial support from the Danish
Cancer Society, the Swedish Research Council, the Novo Nordisk
Foundation (NNF16584), the Danish Council for Independent
Research (13331-00262A), the Lundbeck Foundation, and the
Danish National Research Foundation (Project CARD, DNRF
125). E.W. acknowledges financial support from the German
Research Foundation (CRC1032 and CRC824), Cluster of Excel-
lence NIM, and SinoGerman Center Grant GZ995. S.M.M. ac-
knowledges financial support from the Danish Agency for Science,
Technology, and Innovation (Det Frie Forskningsråd for Teknologi
og Produktion, references 274-08-0534and 12-126894, and Det
Strategiske Forskningsråd, reference 09-065746/DSF) as well as
the International Science and Technology Cooperation of Guang.

REFERENCES1. Al-Dosari, M.S., and Gao, X. (2009). Nonviral gene delivery: principle,
limitations,and recent progress. AAPS J. 11, 671–681.
2. Schlenk, F., Grund, S., and Fischer, D. (2013). Recent developments and perspectives on gene therapy
using synthetic vectors. Ther. Deliv. 4, 95–113.
3. Gao, X., Kim, K.S., and Liu, D. (2007). Nonviral gene delivery: what we know and what is next. AAPS J.
9, E92–E104.
4. Pérez-Martínez, F.C., Guerra, J., Posadas, I., and Ceña, V. (2011). Barriers tonon-viral vector-mediated
gene delivery in the nervous system. Pharm. Res.28, 1843–1858.
5. Nayerossadat, N., Maedeh, T., and Ali, P.A. (2012). Viral and nonviral delivery systems for gene
delivery. Adv. Biomed. Res. 1, 27.
6. Thomas, C.E., Ehrhardt, A., and Kay, M.A. (2003). Progress and problems with the use of viral vectors
for gene therapy. Nat. Rev. Genet. 4, 346–358.
7. Check, E. (2002). A tragic setback. Nature 420, 116–118.
8. Cho, Y.W., Kim, J.D., and Park, K. (2003). Polycation gene delivery systems: escape from endosomes to
cytosol. J. Pharm. Pharmacol. 55, 721–734.
9. Davis, M.E. (2002). Non-viral gene delivery systems. Curr. Opin. Biotechnol. 13,128–131.
10. Lächelt, U., and Wagner, E. (2015). Nucleic Acid Therapeutics Using Polyplexes: A Journey of 50 Years
(and Beyond). Chem. Rev. 115, 11043–11078.
11. Mintzer, M.A., and Simanek, E.E. (2009). Nonviral vectors for gene delivery. ChemRev. 109, 259–302.
12. Pack, D.W., Hoffman, A.S., Pun, S., and Stayton, P.S. (2005). Design and develop-
ment of polymers for gene delivery. Nat. Rev. Drug Discov. 4, 581–593.
13. Boussif, O., Lezoualc’h, F., Zanta, M.A., Mergny, M.D., Scherman, D., Demeneix, B.and Behr, J.P.
(1995). A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo:
polyethylenimine. Proc. Natl. Acad. Sci. USA 92, 7297–7301.
14. Neu, M., Fischer, D., and Kissel, T. (2005). Recent advances in rational gene transfer vector design
based on poly(ethylene imine) and its derivatives. J. Gene Med. 7, 992–1009.
15. Bieber, T., Meissner, W., Kostin, S., Niemann, A., and Elsasser, H.P. (2002).
Intracellular route and transcriptional competence of polyethylenimine-DNA com-
plexes. J. Control. Release 82, 441–454.
16. Wagner, E. (2012). Polymers for siRNA delivery: inspired by viruses to be targeted, dynamic, and
precise. Acc. Chem. Res. 45, 1005–1013.
17. Zheng, M., Librizzi, D., Kılıç, A., Liu, Y., Renz, H., Merkel, O.M., and Kissel, T.(2012). Enhancing in vivo
circulation and siRNA delivery with biodegradable polyethylenimine-graft-polycaprolactone-block-
poly(ethylene glycol) copolymers.Biomaterials 33, 6551–6558.
18. Intra, J., and Salem, A.K. (2008). Characterization of the transgene expression generated by branched
and linear polyethylenimine-plasmid DNA nanoparticles in vitro and after intraperitoneal injection in
vivo. J. Control. Release 130, 129–138.
19. Wightman, L., Kircheis, R., Rössler, V., Carotta, S., Ruzicka, R., Kursa, M., and Wagner, E. (2001).
Different behavior of branched and linear polyethylenimine for gene delivery in vitro and in vivo. J. Gene
Med. 3, 362–372.
20. Ferrari, S., Moro, E., Pettenazzo, A., Behr, J.P., Zacchello, F., and Scarpa, M. (1997).ExGen 500 is an
efficient vector for gene delivery to lung epithelial cells in vitro and in vivo. Gene Ther. 4, 1100–1106.
Polyplex |Summer Training Report |

RESEARCH METHODOLOGY
Research methodology is the procedure adopted for conducting the research study. Research
methodology should be carefully planned as the accuracy, reliability and adequacy of results depend on
Research methodology followed. It gives researcher a guideline by which he can decide which and
procedures will be applicable to a given problem Moreover it helps in the evaluation of research by others
also. So for the research to be successful purposeful and effective, the researcher should plan the
Research methodology before proceedings the research study. The following should be the aspect, which
should be considered while designing a Research methodology.

RESEARCH DESIGN
Research Design is simply the framework or plan for a study that is used as a guide in collecting and
analyzing the data. It is the blue print that is followed in completing a study. As objective of the
research is descriptive in for, the research design must be made accordingly:

➢ Formulating objective of the study.

➢ Designing the method of data collection.
➢ Selecting the sample size.
➢ Collection of data.
➢ Analysis & findings.
➢ Conclusion and interpretations.
➢ Limitations.
➢ Suggestions and recommendations.
Research includes observations and facts used to find enquiries of different kind. Statistical methods
are used in this project and used to infer the results thereof.
Polyplex |Summer Training Report |

SAMPLE DESIGN

➢Type of Universe: - The type of universe selected in sample design.

by researcher is finite to Noida city.

➢ Sampling Unit: - Noida city.

➢ Sampling Elements: - Polyplex Polythene users

➢ Sample Size: - The sample size taken here and covered in this
Project is of 50 respondents

➢Type of Research : Descriptive Research Design

➢Method of Research : Data Collection Method

➢Sampling Method : Convenience Sampling

Polyplex |Summer Training Report |

RESEARCH INSTRUMENTS OR TOOLS USED

Questionnaire: Questionnaire includes a set of questions which are relating to the study objective.
It is used to collect primary data for the purpose of finding solution to the research problem.

DATA COLLECTION

PRIMARY DATA COLLECTION:

Data for the project is collected from the primary sources the questionnaire was filled both by the
customers and retailers. Method of informal interview was also used for the purpose of data
collection.
Polyplex |Summer Training Report |

DATA ANALYSIS AND INTERPRETATION

(1) Do you use material handling plastics Polythene?

Response No. of respondent Percentage (%)

Yes 40 80
No 10 20

Graph
Responce
0

20%

Yes
No
80%

Interpretation
The given chart and table show the no. of respondents who use material handling
plastics Polythene.

It was found that 80% of the respondents like to use material handling plastics
Polythene and 20% of respondents do not like to use plastics Polythene.

Those respondents who not using material handling Polythene are using metal
box. wooden box, plastic tray, Jute & Plastic Bags and etc.
Polyplex |Summer Training Report |

(2) Which Brand of Polythene do you prefer?

Name of Company No. of User Percentage (%)

Polyplex 20 40
Prince 13 26
Supreme 08 16
Syntax 07 14
Other 02 4

PREFERENCE
Polyplex Prince Supreme Syntax other
4%

14%
40%
16%

26%

Interpretation :

The given table and charts shows the brand preference of respondents in plastic
Polythene.

It was found that 40% of total respondents Prefer Polyplex which is at the top and
the second preference of respondents is prince with 26% and third Supreme with
16%.
Polyplex |Summer Training Report |

Response X X-x D2
Polyplex 20 20-10=10 100
Prince 13 13-10=3 9
Supreme 8 8-10=-2 4
Syntex 7 7-10=-3 9
Others 2 2-10=-8 64
∑x=50 0 ∑ D2=186

From arithmetic mean

X=∑X/N =50/5 =10

SD= √∑D2/N = √186/5 = 6.09

Coefficient of SD = SD/mean = 6.09/10 = 0.609

Polyplex |Summer Training Report |

(3) How much Polythene do you use in a day

Response No. of respondents Percentage %

Less than 100 ( mtrs) 41 82
Between 100-500( mtrs) 07 14
More than 500( mtrs) 02 4

No. of respondents

4%
14%

Less than 100 ( mtrs)

Between 100-500( mtrs)
More than 500( mtrs)

82%

Interpretation

The given table & charts shows that the total no. of Polythene used by
respondents.

It was found that our 82% respondent’s majority of them are using less than 100
Polythene, 14 % of them use between 100 to 500Polythene and 4% of them use
more than 500 Polythene. Thus the majority of fever users are more than the
more users in the city of Noida.
Polyplex |Summer Training Report |

(4) Which model of Polythene do you mostly prefer ?

Response No. of respondents Percentage %

Jumbo 12 24
Multi Purpose 23 46
Bottling polythene 05 10
Ice boxes 06 12
FPO Polythene 04 8

No. of respondents
Jumbo Multi Purpose FPO Crates
Ice boxes Bottling crates
10%
12%
24%
8%
46%

Interpretation

The given table and charts shows the percentage of a various model of Polythene
use by the respondent.

If was found that majority of respondent is 46% of the total respondent prefer
multipurpose Polythene. The second position goes to Jumbo Polythene with 24%
third position goes for ice boxes with 12%. Fourth position goes for FPO Polythene
with 10% and users of other Polythene is 8% share .thus it is clear that majority of
user prefer Jumbo and Multipurpose Polythene.
Polyplex |Summer Training Report |

Response X X-x D2
Jumbo 12 2 4
Multipurpose 23 13 169
Botling polythene 5 -5 25
Ice box 6 -4 16
FPO 4 -6 36
∑X= 50 0 ∑D2=250

x=∑X/N = 50/5 = 10

SD = √∑D2//N = √250/5 = 7.07

Coefficient of SD = SD/mean = 7.07/10 = 0.707

Polyplex |Summer Training Report |

(5) What is the frequency of your purchase?

Response No. of respondents Percentage %

Monthly 07 14
Quarterly 16 32
Half Yearly 17 34
Yearly 10 20

Yearly Monthly
20% 14%

Quarterly
Half Yearly 32%
34%

Interpretation: The given table and charts shows how frequently the respondent
purchase the Polythene.

It was found that Polythene out of 100% respondent majority of them, They
prefer Half yearly purchase of Polythene 34% , there after Quarterly 32 %, yearly
20% and Monthly 14% respectively.
Polyplex |Summer Training Report |

X X-x D2
Monthly 07 -5.5 30.25
Quarterly 16 3.5 12.25
Half-yearly 17 4.5 20.25
Yearly 10 -2.5 6.25
∑X= 50 0 ∑D2= 69

x=∑X/N = 50/4 = 12.5

SD= √∑D2/N = √69/4 = 4.15

Coefficient of SD = SD/mean =4.15/12.5 = 0.332

Polyplex |Summer Training Report |

(6) On what basis do you purchase the Polythene?

Response No. of respondents Percentage %

Brand Image 12 24
Quality 23 46
Price 09 18
Others 06 12

Sales

Others
12%
Brand Image
Price
24%
18%

Quality
46%

Interpretation:

The given table and charts shows the basis for the purchase the Polythene.

It was found that majority of people is 46% prefer Quality based purchase
Polythene. The second basis goes for Brand image with 24% and third 18 % and
12% is price base and other basis purchase of Polythene.
Polyplex |Summer Training Report |

X X-x D2
Brand image 12 -0.5 0.25
Quality 23 10.5 110.25
Price 9 -3.5 12.25
Others 6 -6.5 42.25
∑X=50 0 ∑ D2 = 165

x=∑X/N= 50/4 =12.5

SD= √∑D2/N = √ 165/4 = 6.42

Coefficient of SD = SD/mean =6.42/12.5 = 0.51

Polyplex |Summer Training Report |

(7) Does price Effect your buying behaviour?

Response No. of respondents Percentage %

Yes 44 88
No 06 12

Percentage %

12%

Yes
No

88%

Interpretation

The given table & chart shows the effect of price of Polythene on buying behavior
of customer.

It was found that the outcome was a single response. Among 100 % of
respondents, majority of them supported that the price of the Polythene has an
affect buying behavior.

(8) Who take the purchase decision of Polythene?

Polyplex |Summer Training Report |

Response No. of respondents Percentage %

Owner 30 60
Manager 12 24
Store Incharge 06 12
Others 02 4

Others
4% Sales
Store incharge
12%

Manager
24% Owner
60%

Interpretation:

The given table & charts shows the person who take the purchase decision for
Polythene.

It was found that majority of times 60% of total respondent says that owner of
the enterprise takes the purchase decision for Polythene. 24% respondents say
that manager of the enterprise takes purchase decision for Polythene. 12% of
total respondents say that store incharge take the purchase decision for
Polythene. 4% says that other person take the purchase decision.
Polyplex |Summer Training Report |

X X-x D2
Owner 30 17.5 306.25
Manager 12 -0.5 0.25
Store in charge 6 -6.5 42.25
Others 2 -10.5 110.25
∑X=50 0 ∑D2=459

x=∑X/N= 50/4 =12.5

SD= √∑D2/N = √459/4 = 10.71

Coefficient of SD = SD/mean = 10.71/12.5 = 0.856

Polyplex |Summer Training Report |

Are you satisfied with Polythene of polyplex ?

Response No. of respondents Percentage %

Yes 38 76
No 12 24

Percentage %

24%

Yes
No

76%

Interpretation

The given table and charts shows that the satisfaction level of the consumer with
performance of Polythene.

It was found that the majority of the respondent i.e. 76% of the total respondent
is satisfied with the performance of their Polythene.

While only 24% of the consumer were not satisfied with the performance of their
Polythene.
Polyplex |Summer Training Report |

(9) From which medium you aware about the Polythene of polyplex ?

Response No. of respondents Percentage %

By Distributor 08 16
By Agent of Company 15 30
By Friends 06 12
Advertisement 21 42

45

40

35

30

25
No. of respondents
20
Percentage %
15

10

0
By Distributor By Agent of By Friends Advertisement
Company

Interpretation

The given table and charts shows the source of awareness of consumer about the
Polythene. It was found that 42% of total respondent get aware about the
different model of Polythene through advertisement, 30% of them get aware
through agent of the company and 16% aware by distributor and 16% aware from
friends.
Polyplex |Summer Training Report |

X X=x D2
By distributor 8 -4.5 20.25
By agent of 15 2.5 6.25
company
By friends 6 -6.5 42.25
By advertisement 21 8.5 72.25
∑X= 50 0 ∑D2=141

x=∑X/N =50/4=12.5

SD= √∑D2/N = √141/4 =5.93

Coefficient of SD = SD/mean =5.93/12.5 = 0.47

Polyplex |Summer Training Report |

(10) Should company focus on Distribution channel of Polyplex

product?

Response No. of Response Percentage %

Yes 38 76
No 12 24

Percentage %

24%

Yes
No

76%

Interpretation

The given table and charts shows the distribution channel of polyplex product
Polythene.

It was found that the majority of 76 % respondent are agreeing that its
distribution channel and 24% respondents want to improve its and sales
promotion scheme for Polythene.
Polyplex |Summer Training Report |

FINDINGS & SUGGESTIONS

• Majority of user 82% of total user of Polythene, they use less then 100
Polythene.
• Majority user prefers Multipurpose & Jumbo Polythene with 46% &
24%
• The majority of user Polythene 34% prefer half yearly purchase of
Polythene and 32% prefer quarterly.
• Majority of people prefer Price as basis for purchase with 88%.
• Majority of customer are price affecting buying behavior of
Polythene.
• Majority of 60% of total purchase decision taken by owner. I
• Majority of respondent 76% are satisfying with the performance of
Polythene.
• More need of Advertisement and Sales promotion schemes of
Polythene and bins.
Polyplex |Summer Training Report |

SWOT ANALYSIS
A scan of the internal and external environment is an important part of the strategic planning process.

Environmental factors internal to the firm usually can be classified as strengths (S) or weaknesses (W),

and those external to the firm can be classified as opportunities (O) or threats (T). such an analysis of the

strategic environment is refer to as SWOT analysis.

The SWOT analysis provide information that is helpful in matching the firm’s resources and

capabilities to the competitive environment in which it operates, as such it is instrumental in strategy

formulation and selection.

SWOT analysis is very effective way of identifying the strengths and weaknesses, and of examining the

opportunities and threats the organization face. Carrying out an analysis using the SWOT framework helps

us to focus the activities into areas where we are strong and where the greatest opportunities lies. SWOT

analysis is just one of many good techniques that can help us build a competitive position for the

organization.

Definitions

Strengths: - a company’s strength is a successful application of a competency or exploitation of a critical

factors to develop company competitiveness.

Weaknesses: - a company’s weakness is an unsuccessful application of a competency or the non-

exploitation of a critical factor that diminishes company competitiveness.

Polyplex |Summer Training Report |

Opportunities: - an opportunity is an external condition that could positively impact the company’s critical

performance parameters and improve the competitive advantage provided positive action is taken in

time.

Threats: - a threat is an external condition that could negatively impact the company’s critical

performance parameters and diminish competitive advantage provided positive action is not taken in

time.
Polyplex |Summer Training Report |

SUGGESTIONS & RECOMMENDATIONS

The brand image survey brought out various issues, which were supposed to be dealt with, in order to

improve the satisfaction level of customer and retailer. Based on our data collection observation and

analysis we had proposed few suggestions to improve the Brand Image of Polyplex:

1 The customer care services should be improved.

2 Promote TV advertisement :

Polyplex. should promote its scheme by giving advertisement on television. Company should

promote good and heartfelt Slogans and Jingles.

3 Promote print add:

Polyplex. should promote its schemes by giving advertisement in news papers specially VAS

so that the people who dealt with it came to know that benefit of dealing with it.

4 Expand its network:

Polyplex should expand its promotion strategy in rural area. So Network coverage needs to

be expanded in remote areas.

Polyplex |Summer Training Report |

5 Information regarding new plans

Prior information regarding new plans and schemes should be provided to each retailer in time &

promptly as per demand.

6 Among positive side Polyplex has got good services and higher consumer satisfaction

So it should continued this to keep its present customer remain loyal to it.
Polyplex |Summer Training Report |

CONCLUSION

• Polyplex is a customer friendly crate.

• Highly Sophisticated brand, in the perspective of customer.

• Prince and Supreme are growing day to day markets through its prices.

• Price of Polyplex is high comparison to its other competitor.

• Weak distribution channel of Nilkamal.

• It's concerned only major user of Polyplex Product.

Polyplex |Summer Training Report |

LIMITATIONS OF THE STUDY

• Limited time was available for the study, many aspects was left unexplored.

• The area of research was confined to a single city so generalizations cannot be

made for other areas.

• There were few respondents who left certain questions unmarked.

• The study was unfunded therefore the coverage is small.

Polyplex |Summer Training Report |

BIBLIOGRAPHY
1 Kotler , Philip., Marketing Management: Pearson Education

2 Kotler, Philip., and Keller Kevin Lane., Marketing Management: Pearson Education.

3 Kothari,C.R., Research Methodology Methods & Techniques: New Age International (P)Limited,
Publishers.

4 POLYPLEX's Corporate History

5 “REINFORCE”, Polyplex family magazine.

6. www.rcom.co.in
Polyplex |Summer Training Report |

ANNEXURE
Polyplex |Summer Training Report |

Questionnaire

NAME : ________________________________

AGE : ______

CONTACT NO : ________________________

OCCUPATION : ______________________

I - Do you use material handling Polythene?

(a) Yes (b) No

2- Which brand of Polythene do you prefer more?

(a) Polyplex (b) Prince

(c) Supreme (d) Nilkamal

(e) Others

3- How many Polythene do you use in a day?

(a) Less than 100 mtr (b) Between 100 to 500 mtr

(c)More than500 mtr

4- Which model of Polythene do you use?

Polyplex |Summer Training Report |

(a).Tumbo (b) Multipurpose

(c) JBC (d) Ice-Box

(e) Others

5- What is the frequency of your purchase?

(a) Monthly (b) Quarterly

(c) Half Yearly (d) Yearly

6-On what basis do you purchase the Polythene of Polyplex?

(a) Brand Image (b) Quality

(c) Price (D) Other

7-Does Price affect buying behavior?

(a) Yes (b) No

(8) Who take the purchase decision for the Polythene?

Polyplex |Summer Training Report |

(a) Owner (b) Manager (c) In

charge (d)Other

9- Are you satisfied with Polyplex Polythene?

(a) Yes (b) No

10- From which medium you aware about Polythene?

(a) By Advertisement (b) By Distributor

(c) By Sales Agents (d) By Friends

11- Should Company focus on Distribution of channel of Polyplex product?

(a) Yes (b) No

THANK YOU