Introduction: are antibody-producing are fused with mouse myeloma

Introduction:
Cells called hybridomas manufacture antibody in huge amount which is desirable and create monoclonal antibodies.The mouse spleen cells that are antibody-producing are fused with mouse myeloma cells.The obtained secreted antibodies which show specificity are determined by parent spleen cell and the quantity properties are dictated by a myeloma.Here, some chromosomes are lost and some characters of each of the parental cells are preserved in the hybrids.Two scientists, Kohler and Milstein uncovered the hybridoma technology in 1975. this technology introduces monoclonal antibodies in specific cells. Monoclonal antibodies are beneficial for study of parasites antigens. They are used in inhibiting,analysis and managing disease. Cancer antigens,alone or coupled with anti cancer agents are indicated by monoclonal antibodies. They are also used in saving organ transplantation where rejection of organ can occur.

Technique involved in hybridoma technology:
1. Prerequisites to starting a hybridoma project-
– Sterility must be maintained in cabinet where hybridoma production will be introduced .
– An incubator is required in which temperature is controlled, humidity is maintained and gas concentration is maintained. -Liquid nitrogen storage should be facilitated as it is essential for hybridomas maintainment in a low temperature store for cryopreservation.
-Ease of animal holding,aseptic surgical equipment for mouse dissection, water baths of temperature at 37 and 56 degree celcius, centrifugation machine , tissue culture ware are also needed.
2. Materials and media:
-Cell culture medium is used. RPMI 1640 is used,. In addition to it, FCS 10%, Glutamine 2mM, Penicillin 100 IU/ml and streptomycin 100 microgram/ml are used.
– HAT selective medium components ( hypoxanthine, aminopterin, thymidine) are used.
-polyethylene glycol are used.
3. Choice of antigens and immunization schedules:
-Highly specific antibodies against impure antigens are obtained and antibodies can be used to clarify the antigens.
-Spleen cells are used in the antibody-producing hybrids.
-Increasing number of cells in blast transformation ,and depleting insignificant cells are beneficial.
-The ratio of hybrid specific for aimed antigen is enhanced by using purify antigens.
-BALB/c mice strain are used cause myeloma cells set for fusion are of origin of BALB/c.
4. Choice of parent myeloma lines:
-It`s growth should be better and high concentrations of immunoglobulin must be secreted from it. Hybrids must be delayed from parent cells.
-HAT selective medium is required. The main biosynthesis pathway which cause nucleic acid synthesis is blocked by aminopterin . Cell`s multiplications are continued by using salvage pathway in the presence of hypoxanthine and thymidine . If an enzyme needed in the salvage pathway lack in a mutant cell,growth of cell is impossible. Generally, myeloma cells which lack hypoxanthine guanine phosphoribosyl transferase ( HGPRT) are applied. The HGPRT from spleen cell parent is present in fusion product and multiplication occurs in HAT medium.
5. Hybridoma colony and antibody production:
Basic outline of hybridoma technology is following:
A specific antigen is injected into mouse , the antigen-specific plasma cells are produced from the mouse `s spleen and myeloma, a cancerous cell is fused with this cells. This is called hybridoma . The hybrid cell is thus duplicated and innumerable identical daughter clones are obtained and then secretion results in immune cells. These antibodies are called monoclonal antibodies as they produce from only one type of cell. Then, monoclonal antibody is prepared by HAT medium ( hypoxanthine administering thymidine) . To obtain desire antibody against specific antigen, mice are exposed to that antigen.Isolation of splenocytes from the mammal, fusion the B cells with deathless myeloma cells that lack HGPRT gene occur. Polyethylene glycol or Sendai virus helps in fusion. Incubation of fused cells in the HAT medium cause aminopterin in the myeloma cells death as nucleotide inhibition occur. So, death of myeloma cell occur in culture that lack HGPRT enzyme. Death of unfused B cells happen for shorter life span. Only survival portions are B cell-myeloma hybrids because of functional HGPRT gene originated from the B cells. Thus, antibodies are produced by these cells ( characteristics of B cells) and cells are immortal ( characteristics of myeloma cells). Then, dilution of incubated medium into multi well plates are done so that only one cell can be present in each well and checking for desired antibody are taken place.The antibodies in a all came from the same B cell ,will be in the direction of same epitope and so, this is called monoclonal antibodies. Now, a hybrid oma colony is created ,continuous growth takes place in culture medium and antibody is obtained.

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6.Rapid primary screening process:
Hybridomas that create antibodies of appropriate specificity is identified and selected only. Secondary enzyme that is labelled with conjugate , hybridoma that culture supernatant and chromatogenic substrate can be breed and a colored product is formed. it indicates a positive hybridoma. Alternatively, ELISA can be done. Here, adsorbtion of antigen to the the bottom of 96-well plates,incubation for growth of hybridomas occur. The desired antibody in the sample remain bound to antigen and is detected by an immunoconjugate. Immunoconjugate is consists of two components, one antibody is specific for an epitope that remains in the constant domain in the first antibody. It acts as anti-antibody.Second one is alkaline phosphatase enzyme. Immunoconjugate is retained in the well during the immobilization at first incubation of antibody. After washing colorless substrate of enzyme ( ex. p-nitrophenyl phosphate) is converted to a colored product ( ex. p-nitrophenol) by the enzyme( ex. alkaline phosphatase). After incubation and the termination of enzyme function, ELISA reader quantify the optical density of product.

7. Cloning:
After screening , cloning can be done by three different tenchniques( cloning by technique of limiting dilutions,cloning using semi-solid agars and cloning and selection using the fluorescence-activated cell sorter).
8. Cryopreservation:
It is essential against the loss of beneficial lines.Hybridoma should be freezed down as soon as possible to reduce chromosome loss.
9. Characterization of monoclonal antibodies:
Monoclonal antibodies should be characterized with respect to following parameter:
-specificity is determined for specific antigen.
-titer: By chosing for screening and concluding the highest dilution at which a positive outcome can be achieved.
-affinity of binding can be measured.
-storage and stabilty are determined to know the effect of manipulations,such as lyophilization.
-Immunoglobulin class/subclass: For recognition of IgG1, igG2a, IgG2b,IgG3 and IgM, sets of antisera are used.
-monoclonality:only one subclass of antibody/ only one cell type should be present in hybridoma.
10. Production,purification and labelling of monoclonal antibodies:
Monoclonal antibody can be manufactured by two processes ( tissue culture or by maturing the hybridoma as a tumor in mice). Then, ammonium sulfate can precipitate antibody and ion-exchanging chromatography and antigen affinity chromatography can be used for higher extend of antibody purity. At last, Labelling of monoclonal antibodies can be achieved by radioistopic labelling or by fluorescence labelling.

Introduction:
In this project, I am going to describe about my new experience of learning, how to get a domain name, design a website using Wix and publish it online to interpret in today’s business world.

Website Address:
https://tangypizzacom.wixsite.com/0000

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Type of website:
It was a wonderful experience of building a stunning website with Wix.com. The type of website I have designed is a Business Website and this business website will be selling and supporting a pizza business located at 12660 68A, Ave Surrey BC V3W1H3. The name of my business is TANGY PIZZA.
On my business website you will find different attractive templates
? Pizza Business selected suitable template for my website
? Header menu
? Logo and Tag Line
? Backgrounds
? Images and video
? Detailed menu
? Online ordering facility
? Make a Reservation for party
? Location including maps
? Social media connectivity
? Contact info (CTA)
Website Purpose:
As it is a business website and in today’s world of advance technology it is very difficult to run and make your brand hold a position in today’s market by using online facilities such as designing and using a business website. The main purpose of my business website is to increase brand awareness, drive traffic towards my business website, grow revenue by increasing signups or sales, boost brand engagement and provide effective customer service.

Website Design:
The basic design principle of my business website is to accomplish the main purpose of designing my website. Communication is very important to run business to accomplish it. I have used Time New Roman font style in the major parts of website and in description I have used Arial font style for the better understanding of my business customers and visitors. These typefaces are very eye Catchy and attention seeking.
I have used very basic colours in web designing such as white, black, gray that provides relaxation to eyes while reading and also, I have used very attractive and high-quality images and a video that describes the personality and quality of my business products.
Design Plan:
Website’s Goals and Target Audience:
Goals:
I. Increase brand awareness: It’s important to stay relevant. If customers aren’t aware of my brand, the business won’t survive. By using social-media platform customers will become aware of my brand and interact with it on different social media platforms, they are more likely to recommend it to their friends by liking posts on Facebook and retweeting on Twitter.
II. Drive traffic in my business website: Traffic is important to all sites, especially for online selling practice. By using social-media platform I will influence more and more traffic on my official business website.
III. Grow revenue by increasing signups or sales: I will increase my sales and signups by using social-media platforms and advertising on them. By doing this, I will influence my business network and increase brand awareness in consumers and gain word of mouth that will be my biggest asset for my business success.

IV. Boost brand engagement: Engagement is the most important reason why we all use social-media. By using different social-media platforms I will improve my brand perception, loyalty, and word of mouth recommendations.

V. Effective customer service: Having a good customer service on social media can help to increase revenue, customer satisfaction score, and retention.

Target Audience:
The target market for the Tangy Pizza would be a consumer that are of the age of 16-44. I am targeting both male and females for the breakfast. That would give us a huge potential target market of customers in Surrey. The reason we chose this target market was because of the small sample size survey that I have practiced.

The results showed that 95% were between the ages of 18-21, 3% from the ages of 22-25, and 2% age 30 plus. The total number of people surveyed was 43 people. Due to a small university sample size I have used the results as directional and predicted the age range of 16-44. We would also be targeting families that have an average household income. (Shah, 2018)

Products, Services: Tangy pizza is product and service-based enterprise. This business is well prepared for providing high quality products such as different kinds of pizza’s and delicious variety of pasta and beverages to make customer feel more joyful. Tangy Pizza promise to provide its fresh and crisp high quality of pizza & pasta

Website Structure:

Website General Content
? Home page
? Menu
? Order online
? Reservations
? About us
? Locate
? Images & Multimedia

Website’s Navigation System
It is quite clear by looking at the design scheme of my business website. I have designed my website in such a way that everyone can access my business website with ease and decide what he or she is looking to order accordingly.
For better understanding I have used TIMES NEW ROMAN font in my website and for description of my company’s product menu I have used ARIAL fort. According to my research and understanding I feel that the above-mentioned fonts are very reliable in terms of reading a text document on screen.

Design the look and feel of the website
In the process of designing my business website. Wix proved to be very helpful to me, loaded with numerous options and features and it is very user friendly.

Test, Publish, and Maintenance of website
Testing publishing and maintaining of your website is very convenient on Wix.com.
The movement I finished designing my business website, using Wix I published my website by clicking on the top right Blue button and tested weather the design and menu features and several other facilities are working in order as I have planned.
Maintenance is very important part of everything either it is a small business or a big business website because, every business learns from its mistakes. Maintenance is very easy in Wix platform and one can easily maintain it by using its different facilities such as tracking and analytical.
Website Linkage with Social Media Platforms
I am using 3 major social-media platforms for the promotion and brand awareness of my business. These three social-media platforms are as follows:
1. Facebook
2. Tweeter
3. Instagram
Conclusion
It is evident, that after performing above describes web designing project I am now capable of creating my domain name, design a website and publishing of website for further business purposes and make my business available online to interact in today’s business world.

INTRODUCTION.
In the ever changing world of business it is important that we take into cognizance the role that our behavioral and entrepreneurial pattern have in each other’s lives and our ability to fully live out our individual rights. It is because of the neglect for such that the United Nations Global Compact Initiative has human rights as one of its core principles. This piece of work is going to be about the benefits and challenges for different international business in relation to the adoption of the global compact, focusing on human rights. The Global Compact as defined by Suder (2008) refers to the interaction of UN agencies, business, non-governmental organizations, societies and governments in an effort to build a sustainable international business environment. Buzan and Lawson (2015) add onto this definition by describing the Global Compact as an interactive approach to achieving responsible and sustainable business. As one of the principles of the Global Compact Initiative, human rights are privileges and freedom which every person is entitled to by virtue of being human. All basic human rights are found in the United Nations Universal Declaration of Human Rights. They include the right to life, the right to freedom of association, the right to freedom of speech, the right to choice etc. Brems (2001) adds onto this definition by describing human rights as principles and rights given to all people by virtue of being human in an attempt to safeguard their being and are based on ideas of fairness and equality. In this essay we will be assessing some of the major human rights issues and why they are of a global concern and how a global, interactive approach to their protection is the best way forward.

HUMAN RIGHTS ISSUES IN THE WORKPLACE.
There are several human rights issues that take place in the work place, a lot of them intentional and others unintentional. In this segment we assess the three major likely human rights abuses in the workplace which include discrimination, child labor and working in high risk environments. There are however other human rights issues like disregard for employee rights in cyber space, women and minority rights etc.
WORK PLACE DISCRIMINATION.
Chin (2009) defines workplace discrimination as the unfair and prejudicial treatment of certain individuals or groups of people in the workplace due to prejudice. Discrimination comes in two forms, it can be direct or indirect. Direct discrimination is the actual unfair treatment of other individuals whilst indirect discrimination occurs through the passing of rules and regulations which sideline other people in the work place. People discriminate against others for several reasons including sex, race, disability, nationality etc. Gregory (2003) argues that marginalized groups in the workplace are put at a disadvantage as their ability to actualize is limited to how they are viewed by those discriminate them. This has a negative repercussion in terms of establishing individual identities but could also harm motivation and productivity in the place of work.
CHILD LABOR.
With an increase in the utilization of child labor globally it is also important that we take into consideration children’s rights in such working environments. Child labor can be defined as the exploitative and often illegal employment of children to work in industrial sectors, often manufacturing and agricultural sectors. It is disadvantageous for a variety of reasons, amongst which is the exploitation of children in the workplace by making them work long hours with little and unguaranteed pay. The other is that children are often exposed to things like trafficking and abuse by employers’ who often disregard their most basic rights. But just as important is the need to give children access to things like a decent home, basic education and access to clean water and food, which rarely occurs when young children are turned into laborers. Hindman (2014) describes child labor as a global pandemic which occurs in both the developed and under developed world and an issue we should all pro-actively stand against.
WORK IN HIGH RISK ENVIRONMENTS.
Being employed in a mine, in a high tech institution or in a politically volatile area all constitute working in high risk environments. Different businesses have different probabilities of risk attached to them but the mere existence of this risk is reason enough to motivate the global communicate to mitigate against any such risk. This is important especially in the world we live in today where unemployment is rife and individuals do all that they can even when it put them in danger just in order for them to be able to make a decent living. According to Francioni (2001) the people most likely to be affected by human rights abuses in high risk environments are those without any realistic opportunities and are susceptible to abuse by those with the authority, they include migrant workers, women and children.

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SAFEGUARDING HUMAN RIGHTS .
When taking into consideration the abuses that occur in the workplace, we also need to look at the possible solutions to curbing these problems and how corporations can proactively participate in ensuring this. We are going to be discussing employee education, avoiding corporate complacency and attaching corporate liability to human rights abuses.
EMPLOYEE EDUCATION.
Adopting a human rights training plan can help staff to understand and respect one another in workplaces. A lot of our prejudices are things we are capable of unlearning but the process of unlearning is one that needs to accommodate the fact that we need time and some degree of external influence to recognize wrong in some of our behavioral patterns. The same can be same for violations that occur subconsciously and without ill intent, by training our staff we allow them to be able to pick such things out and avoid them in advance. Colella and King (2018) argue that in order to have a tolerant working environment a careful consideration of factors like disability and advancing issues of identity should be prioritized as fields we all need to understand.
AVOIDING CORPORATE COMPLACENCY.
Corporations need to do more than just train their employees in order to achieve human rights, they also have the added responsibility to make policy and laws which achieve this. An equity and human rights policy is important in ensuring the protection of human rights. For example, the establishment of an equity policy for the equal treatment of women or people with disabilities in the workplace would allow these two minority groups to become more comfortable in the workplace. Wilson (2013) argues that an equitable environment allows for individual to excel in their fields which will yield tremendous fruits for the business.
CORPORATE LIABILITY FOR HUMAN RIGHTS ABUSES.
The government has the greatest responsibility to ensure our human rights are adhered to and to avoid any abuses that could occur if this doesn’t happen. It is in consideration of this tat governments should also take action against corporations which actively undermine and disregard our human rights. The State has a duty to enact laws to protect its population from human rights abuses and ensure that companies and individuals within these companies don’t act in any way that contravenes these laws. Amongst others could be the enactment of consumer legislation, workplace safety laws and humanitarian laws to ensure maximum safeguard of human rights. Joseph (2004) argues that attaching corporate liability for human rights violations is important in order to achieve consistent jurisprudence and that the criminalization of human rights abuses would help clearly identify the obligation of corporations, their staff and victims in creating and sustaining a more equitable environment.

CONCLUSION.
Human rights are an essential part of our existence are have been put in place no only to protect us but to ensure that we have equal access to opportunities around us. It is important that in all parts of the worlds these be adhered to so as to set the principle of their significance and to maximize the value of human life wherever we are. It is also important to protect the rights of those most vulnerable amongst us, these include women, children, the disabled and migrants in our countries. By so doing not only do individuals benefit but so do corporations because the productivity levels of people that are well taken care of is significantly higher than those who are not. Our ability to protect the most basic human rights could extend to flourishing industries and eventually flourishing states.

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